KR102154880B1 - Pharmaceutical nanoparticles showing improved mucosal transport - Google Patents

Abstract

Compositions with improved particle transport in mucus are provided. The composition comprises modifying the surface coating of the particles comprising a drug having a low water solubility. In some embodiments, the surface coating comprises a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer, such as poly(vinyl alcohol) (PVA). Using these compositions and methods, efficient transport of particles of a drug through the mucous barrier in the body can be achieved for a wide range of applications, including drug delivery, imaging and diagnostic applications.

Description

Pharmaceutical nanoparticles showing improved mucosal transport {PHARMACEUTICAL NANOPARTICLES SHOWING IMPROVED MUCOSAL TRANSPORT}

The present invention relates generally to particles, compositions and methods that aid in particle transport in mucus.

The mucus layer present at various points of entry into the body, including the eyes, nose, lungs, gastrointestinal tract and female reproductive organs, is naturally sticky, effectively traps pathogens, allergens and debris and facilitates mucus turnover. It serves to protect the body against them by rapidly removing them. For effective delivery of therapeutic, diagnostic or imaging particles through the mucous membrane, the particles must easily penetrate the mucous layer to avoid mucus adhesion and rapid mucus clearance. Some types of evidence suggest that conventional nanoparticles cannot cross the mucosal barrier. However, polymeric nanoparticles (either covalently or non-covalently) modified (covalently or non-covalently) with special surface coatings have recently been shown to be able to diffuse in physiologically rich mucus samples as quickly as they diffuse in water. Proved in. These polymer-based mucus-penetrating particles (MPP) can encapsulate various therapeutic agents, imaging agents, or diagnostic agents to enable drug delivery, diagnosis, or imaging applications.

Despite these improvements, very few surface coatings have been found to promote particle penetration into mucus. Thus, improvements in compositions and methods comprising mucus-penetrating particles for delivery of pharmaceutical agents may be beneficial.

Summary of the invention

The present description generally relates to particles, compositions and methods that aid in particle transport in mucus. In some embodiments, the compositions and methods include synthetic polymers (eg, poly(vinyl alcohol), PVA) having pendant hydroxyl groups on the backbone of the polymer, which promote mucus penetration. The subject matter of this application includes, in some cases, interrelated products, alternative solutions to certain problems, and/or many different uses of structures and compositions.

In one set of embodiments, a method of forming coated particles is provided. The method includes coating a plurality of core particles with a surface-altering agent to form the coated particles, wherein the surface-altering agent has a molecular weight of about 1 kDa or more and about 1000 kDa or less and is on the backbone. And synthetic polymers having pendant hydroxyl groups, wherein the polymer is hydrolyzed by at least about 30% and less than about 95%. Each of the core particles contains a drug or a salt thereof. The coated particles have a relative velocity of greater than 0.5 in the mucus.

In another set of embodiments, a composition comprising a plurality of coated particles is provided. Each of the coated particles comprises a core particle comprising a drug or a salt thereof, and a coating comprising a surface modifier surrounding the core particle. Surface modifiers include synthetic polymers having a molecular weight of at least about 1 kDa and up to about 1000 kDa and having pendant hydroxyl groups on the backbone, wherein the polymer is hydrolyzed by at least about 30% and less than about 95%. The coated particles have a relative velocity of greater than 0.5 in the mucus.

In another set of embodiments, a method is provided. The method includes delivering a composition comprising a plurality of coated particles to a mucus or mucous membrane. The coated particles include core particles comprising a medicament or a salt thereof, and a coating comprising a surface modifier surrounding the core particles. Surface modifiers include synthetic polymers having a molecular weight of at least about 1 kDa and up to about 1000 kDa and having pendant hydroxyl groups on the backbone, wherein the polymer is hydrolyzed by at least about 30% and less than about 95%. The coated particles have a relative velocity of greater than 0.5 in the mucus.

Other advantages and novel features of the invention will become apparent from the following detailed description of various non-limiting embodiments of the invention when considered in conjunction with the accompanying drawings. In the event that this specification and documents incorporated by reference contain conflicting and/or inconsistent disclosures, the present specification will control. If two or more documents incorporated by reference contain conflicting and/or inconsistent disclosures with respect to each other, the document with the later effective date shall prevail.

Non-limiting embodiments of the present invention will be described by way of example with reference to the accompanying drawings, which are schematic and are not intended to be enlarged to scale. In the figures, each of the illustrated identical or nearly identical components is typically indicated by a single number. For the purpose of clarity, not all components are shown in all drawings, and not all components of each embodiment of the invention are shown unless illustration is essential to enable those skilled in the art to understand the invention. In the drawing,
1 is a schematic diagram of a muco-penetrating particle having a coating and a core of a solid medicament, according to one set of embodiments.
2A is an ensemble averaged velocity <V _average > in human cervicovaginal mucus (CVM) for PSCOO particles coated with various poly(vinyl alcohol) (PVA) according to one set of embodiments. Is a plot showing.
2B is a plot showing the relative velocity <V _mean > _relative in CVM for PSCOO particles coated with various PVAs, according to one set of embodiments.
3 is a plot showing the relative velocity <V _mean > _relative in CVM for PSCOO particles incubated with various PVAs mapped according to the molecular weight and degree of hydrolysis of PVA, according to one set of embodiments. Each data point represents the <V _mean > _relative to a particle stabilized with a particular PVA.
4A-4B are plots showing bulk transport in an in vitro CVM of PSCOO nanoparticles coated with various PVAs, according to one set of embodiments. Negative control is uncoated 200 nm PSCOO particles; Positive control is 200 nm PSCOO particles coated with Pluronic® F127. Figures 4A-4B show data obtained using two different CVM samples.
Figures 5A-5B are poly(lactic acid) (PLA) nanoparticles prepared by emulsification with various PVAs, as measured by multiple-particle tracking in CVM, according to one set of embodiments. It is a plot showing the ensemble-average velocity <V _mean > (Figure 5A) and relative sample rate <V _mean > _relative (Figure 5B) for (sample).
6 is a plot showing the relative velocity <V _average > _relative in CVM for PLA nanoparticles prepared by emulsification using various PVA mapped according to the molecular weight and degree of hydrolysis of PVA, according to one set of embodiments. Each data point represents the <V _mean > _relative to a particle stabilized with a particular PVA. The "+" sign indicates measurements in multiple CVM samples.
Figures 7A-7B show ensemble-average velocity <V _mean > (Figure 7A) and relative for pyrene nanoparticles (sample) and control, as measured by multi-particle tracking in CVM, according to one set of embodiments. Sample rate <V _mean > is a plot showing the _relative (Figure 7B).
Figures 8A-8F are representative for pyrene/nanocrystals (sample = pyrene nanoparticles, positive = 200 nm PS-PEG5K, negative = 200 nm PS-COO) obtained with various surface modifiers, according to one set of embodiments. CVM velocity (V _mean ) distribution histogram.
9 is a plot of relative velocity <V _mean > _relative for pyrene nanocrystals coated with PVA in CVM mapped according to the molecular weight and degree of hydrolysis of PVA, according to one set of embodiments.
10 is a plot showing the pharmacokinetics of loteprednol etabonate (LE) in the cornea of New Zealand white rabbits in vivo. 3 kinds of 1 to 50 μL per dose to each eye of the rabbit LE-MPP (i.e., LE-F127, LE- Tween (Tween) 80, and PVA-LE) for each one of the species or Rotterdam Max (Lotemax) ^® Provided. PVA had a molecular weight of about 2 kDa and was hydrolyzed about 75%. The dose of LE was 0.5% in all cases. Error bars represent standard error of the mean (n = 6).
11 is a plot showing the mucus mobility of loteprednol etabonate (LE) particles coated with various PVAs as a function of the molecular weight (MW (kDa)) and degree of hydrolysis (% hydrolysis) of PVA. Samples that do not form particles within the target range are specified as "not formulated".

details

Particles, compositions and methods are provided that aid in particle transport in mucus. The compositions and methods, in some embodiments, include modifying the surface coating of particles comprising a medicament having low aqueous solubility. In some embodiments, the surface coating comprises a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer, such as poly(vinyl alcohol) (PVA). These compositions and methods can be used to achieve efficient transport of particles of a drug across the mucous barrier in the body for a wide range of applications, including drug delivery, imaging and diagnostic applications. In certain embodiments, pharmaceutical compositions comprising such particles are suitable for routes of administration involving particles that cross the mucosal barrier.

In some embodiments, the compositions and methods include the use of PVA to aid in particle transport in mucus. The compositions and methods may include preparing mucus-penetrating particles (MPPs), for example by an emulsification process in the presence of a specific PVA. In certain embodiments, the compositions and methods include preparing MPPs from pre-fabricated particles by non-covalent coating with specific PVA. In other embodiments, the compositions and methods may include preparing the MPP in the presence of a specific PVA without any polymeric carrier or with minimal use of a polymeric carrier. However, it should be appreciated that in other embodiments, polymeric carriers may be used.

PVA is a water-soluble nonionic synthetic polymer. Due to its surface-active properties, PVA is widely used in the food and pharmaceutical industries as a stabilizer for emulsions and, in particular, to enable the encapsulation of a wide variety of compounds by emulsification techniques. PVA has a status of "Generally Recognized as Safe" or "GRAS" by the Food and Drug Administration (FDA), and is a otic, intramuscular, intraocular, intravitreal, iontophoretic, ophthalmic, oral, topical, and transdermal drug. It is being used in products and/or drug delivery systems.

In certain previous studies, many have described PVA as a mucoadhesive polymer, which suggests or reports that incorporation of PVA in the particle formulation process results in strongly mucoadhesive particles. Surprisingly and contrary to the established opinion that PVA is a mucoadhesive polymer, the inventors believe that compositions and methods utilizing a particular grade of PVA within the context of the present invention aid particle transport in mucus and in certain applications described herein. It was found that it was not adhesive. Specifically, mucus-penetrating particles can be prepared by tailoring the degree of hydrolysis and/or molecular weight of PVA, which was previously unknown. These findings significantly expand the arsenal of techniques and components applicable to prepare MPPs, and advantageously, certain existing substances that have been found to promote mucus penetration, such as poly(ethylene glycol) (PEG) and (poly( Ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymer (Pluronics®) addresses certain limitations.

In particular, in terms of drug delivery applications, there may be certain limitations associated with PEGylated or Pluronic®-coated MPPs: A) PEG-containing blocks for preparing PEGylated or PEGylated MPPs of pre-assembled particles The use of copolymers can create new inert ingredients as prescribed by the FDA. These novel ingredients may require expensive and time-consuming preclinical and clinical safety studies and present significant regulatory challenges by the FDA, which may limit the practical possibilities of PEGylated MPPs. B) It is difficult to directly prepare Pluronic®-coated MPPs through the emulsification technique, the most popular and most common approach to nano- and microencapsulation. This limitation significantly reduces the range of compounds that can be efficiently encapsulated with Pluronic®-coated MPP. In particular, this may make it impossible to efficiently encapsulate water-soluble to weakly water-soluble ingredients (e.g., drugs, proteins, oligonucleotides, etc. with water solubility greater than about 1 mg/mL) that are most commonly encapsulated by double emulsification techniques. have. Thus, improvements in compositions and methods that allow direct preparation of MPPs by emulsification techniques would be beneficial. C) Pluronics® is currently not approved by the FDA for dosage via the auricular, intramuscular, intraocular, intravitreal routes, which are the approved dosage routes for PVA. In addition, the maximum Pluronic® concentration currently approved by the FDA for ophthalmic dosage is 0.2%, while for PVA it is 1.4%. Thus, improvements in the compositions and methods that enable the preparation of MPPs by using PVA (e.g. instead of Pluronics®) greatly increase the clinical development of the mucous-penetrating particle technology as a whole, especially the above-mentioned dosage route. There is a possibility to promote it.

In some embodiments described herein, compositions and methods for making particles, including certain compositions and methods for making particles with increased transport through the mucosal barrier, address one or more or all of the concerns described above. Specifically, in some embodiments, the compositions and methods comprise preparing mucus-penetrating particles by an emulsification process in the presence of certain PVA. Advantageously, by using PVA for MPP production, certain limitations of PEGylated or Pluronic®-coated MPPs can be solved with respect to drug loading, range of encapsulated materials, and/or complexity of clinical development.

While in some embodiments it may be advantageous to form mucous-penetrating particles without the use of PEG or Pluronics®, it should be appreciated that in other embodiments PEG and/or Pluronics® may be included in the compositions and methods described above.

In some embodiments, the particles described herein have a core-shell type arrangement. The core may comprise any suitable material, such as a solid medicament having relatively low water solubility or a salt thereof, a polymeric carrier, a lipid, and/or a protein. In addition, the core may comprise a gel or liquid in some embodiments. The core may be coated with a coating or shell comprising a surface modifier that promotes the mobility of the particles in the mucus. As described in more detail below, in some embodiments the surface modifier may include a polymer (eg, synthetic polymer or natural polymer) having pendant hydroxyl groups on the backbone of the polymer. The molecular weight and/or degree of hydrolysis of the polymer can be selected to impart certain transport characteristics to the particles, such as increased transport through mucus.

Non-limiting examples of particles are now provided. As shown in the exemplary embodiment of FIG. 1, particle 10 comprises a core 16 (which may be in the form of a particle, referred to herein as a core particle) and a coating 20 surrounding the core. do. In one set of embodiments, a significant portion of the core is formed of one or more solid agents (e.g., drugs, therapeutics, diagnostic agents, imaging agents) that can cause certain beneficial and/or therapeutic effects. The core can be, for example, a nanocrystal (ie, nanocrystalline particles) of a drug. In other embodiments, the core may comprise a polymeric carrier, optionally with one or more agents encapsulated or otherwise associated with the core. In still other cases, the core may comprise a lipid, protein, gel, liquid, and/or another suitable material to be delivered to the subject. The core comprises a surface 24 to which one or more surface modifiers can be attached. For example, in some cases, the core 16 is surrounded by a coating 20 comprising an inner surface 28 and an outer surface 32. The coating may be formed, at least in part, of one or more surface modifiers 34, such as polymers (e.g., polymers or synthetic polymers having pendant hydroxyl groups), capable of associated with the surface 24 of the core. I can. Surface modifiers 34 can be, for example, covalently attached to the core particles, non-covalently attached to the core particles, adsorbed to the core, or ionic interactions, hydrophobic and/or hydrophilic interactions, electrostatic interactions, van der It can be associated with the core particles by being attached to the core through a Balse interaction, or a combination thereof. In one set of embodiments, a surface modifier, or portion thereof, is selected to facilitate transport of the particles through the mucosal barrier (eg, mucus or mucous membrane).

Particle 10 may optionally comprise one or more components 40, such as a targeting moiety, protein, nucleic acid, and bioactive agent, which may optionally impart specificity to the particle. For example, a targeting agent or molecule (e.g., a protein, nucleic acid, nucleic acid analog, carbohydrate, or small molecule), if present, can assist in directing the particle to a specific location in the subject's body. The location can be, for example, a tissue, a particular cell type, or an intracellular compartment. One or more components (40), if present, may be associated with the core, coating, or both; For example, they may associate with the surface 24 of the core, the inner surface 28 of the coating, the outer surface 32 of the coating, and/or be embedded in the coating. The one or more components 40 may be associated through covalent bonds, absorption, or attached through ionic interactions, hydrophobic and/or hydrophilic interactions, electrostatic interactions, van der Waals interactions, or combinations thereof. In some embodiments, the component may be attached to (eg, covalently) one or more of the coated particles to the surface modifier using methods known to those of skill in the art.

It is to be understood that components and configurations other than those shown in FIG. 1 or described herein may be suitable for certain particles and compositions, and in some embodiments not all of the components shown in FIG. 1 are necessarily present.

In one set of embodiments, the particle 10, upon introduction into the subject, comprises one or more components in the subject, such as mucus, cells, tissues, organs, particles, fluids (e.g., blood), portions thereof, and combinations thereof. Can interact with In some such embodiments, the coating of the coated particles 10 comprises a surface modifier or other component having properties that enable favorable interaction (e.g., transport, binding, adsorption) with one or more substances from a subject. Can be designed to For example, the coating may contain a surface modifier or other component having a specific hydrophilicity, hydrophobicity, surface charge, functional group, specificity for binding, and/or density, that promotes or reduces certain interactions in the subject. I can. One specific example is by selecting the specific hydrophilicity, hydrophobicity, surface charge, functional group, specificity for binding, and/or density of one or more surface modifiers that reduce the physical and/or chemical interactions between the particles and the mucus of the subject. , To improve the mobility of particles through mucus. Other examples are described in more detail below.

In some embodiments, once the particles are successfully transported across the mucosal barrier (eg, mucous or mucous membrane) in the subject, further interactions between the particles may occur in the subject. Interactions can, in some cases, occur through the coating and/or core, for example, from one or more components of the subject to the particles 10, and/or from the particles 10 to one or more components of the subject. Exchange of substances (eg, pharmaceuticals, therapeutic agents, proteins, peptides, polypeptides, nucleic acids, nutrients). For example, in some embodiments in which the core is formed of or comprises a medicament, the degradation, release and/or transport of the medicament from the particles may cause certain beneficial and/or therapeutic effects in the subject. As such, the particles described herein can be used in the diagnosis, prevention, treatment or management of a particular disease or physical condition.

Specific examples of the use of the particles described herein are provided below in the context of being suitable for administration to the mucosal barrier (eg, mucous or mucous membranes) in a subject. While many embodiments herein are described in this context and in the context of providing benefits to diseases and conditions associated with the transport of substances across the mucosal barrier, the invention is not so limited and the particles, compositions, It should be appreciated that the kits and methods can be used to prevent, treat, or manage other diseases or physical conditions.

Mucus is a viscous, viscoelastic gel that protects against pathogens, toxins and foreign objects at various points of entry into the body, including the eyes, nose, lungs, gastrointestinal tract and female reproductive organs. Many synthetic nanoparticles are strongly mucoadhesive and are effectively captured in the rapidly cleared surrounding mucus layer, which enormously limits their distribution throughout the mucous membrane as well as penetration into the underlying tissue. The residence time of these trapped particles is limited by the rate of replacement of the surrounding mucus layer, which ranges from a few seconds to several hours, depending on the organ. In order to ensure effective delivery of particles comprising a drug (e.g., therapeutic, diagnostic, and/or imaging agent) through the mucous membrane, these particles must diffuse easily through the mucous barrier to avoid mucus adhesion.

It has recently been demonstrated that modifying the surface of polymeric nanoparticles with mucus-penetrating coatings can minimize adhesion to mucus and thus allow rapid particle penetration across mucus barriers. Specifically, polymer nanoparticles with a size of about 500 nm are covalently with dense coatings of low molecular weight PEG (2 kDa -5 kDa) or specific Pluronic ^® molecules (e.g. P103, P105, F127) It has been found that when coated non-covalently with, they are able to penetrate human mucus almost as quickly as they move in pure water and at a rate nearly 100 times faster than similar-sized uncoated polymer particles. Despite these improvements, very few surface coatings have been found to promote the penetration of particles into mucus. Accordingly, improvements in compositions and methods comprising mucus-penetrating particles for delivery of medicaments may be beneficial.

Core particles

As described above with reference to FIG. 1, the particles 10 may include a core 16. The core can be formed from any suitable material, such as organic material, inorganic material, polymer, lipid, protein, or combinations thereof. In one set of embodiments, the core comprises a solid. The solid can be, for example, a crystalline or amorphous solid, such as a crystalline or amorphous solid drug (eg, a therapeutic agent, a diagnostic agent, and/or an imaging agent), or a salt thereof. In other embodiments, the core may comprise a gel or liquid (eg, oil-in-water or water-in-oil emulsion). In some embodiments, more than one agent may be present in the core. Specific examples of medicaments are provided in more detail below.

The medicament may be in any suitable amount, for example, at least about 0.01 wt%, at least about 0.1 wt%, at least about 1 wt%, at least about 5 wt%, at least about 10 wt%, at least about 20 wt%, about 30 wt% of the core. wt% or more, about 40 wt% or more, about 50 wt% or more, about 60 wt% or more, about 70 wt% or more, about 80 wt% or more, about 85 wt% or more, about 90 wt% or more, about 95 wt% Or more or about 99 wt% or more in the core. In one embodiment, the core is formed from 100 wt% of the drug. In some cases, the medicament is about 100 wt% or less, about 90 wt% or less, about 80 wt% or less, about 70 wt% or less, about 60 wt% or less, about 50 wt% or less, about 40 wt% or less, about 30 wt% or less, about 20 wt% or less, about 10 wt% or less, about 5 wt% or less, about 2 wt% or less, or about 1 wt% or less in the core. Combinations of the aforementioned ranges are also possible (eg present in an amount of about 80 wt% or more and about 100 wt% or less). Other ranges are also possible.

In embodiments in which the core particles comprise a relatively large amount of the agent (e.g., at least about 50 wt% of the core particles), the core particles generally have an increased amount of agent compared to particles formed by encapsulating the agent in a polymeric carrier. Has loading This is an advantage for drug delivery applications, as higher drug loading means that fewer particles may be required to achieve the desired effect compared to the use of particles containing a polymeric carrier.

As described herein, in other embodiments in which a relatively large amount of polymer or other material forms the core, a smaller amount of the agent may be present in the core.

The core may be formed from a solid material having a variety of water solubility (i.e., solubility in water, optionally with one or more buffers), and/or a solubility in a solution in which the solid material is coated with a surface modifier. For example, a solid material is about 5 mg/mL or less, about 2 mg/mL or less, about 1 mg/mL or less, about 0.5 mg/mL or less, about 0.1 mg/mL or less, about 0.05 mg/mL at 25°C. Or less, about 0.01 mg/mL or less, about 1 μg/mL, about 0.1 μg/mL or less, about 0.01 μg/mL or less, about 1 ng/mL or less, about 0.1 ng/mL or less, or about 0.01 ng/mL or less It may have water solubility (or solubility in the coating solution). In some embodiments, the solid material is at least about 1 pg/mL, at least about 10 pg/mL, at least about 0.1 ng/mL, at least about 1 ng/mL, at least about 10 ng/mL, at least about 0.1 μg/mL, About 1 μg/mL or more, about 5 μg/mL or more, about 0.01 mg/mL or more, about 0.05 mg/mL or more, about 0.1 mg/mL or more, about 0.5 mg/mL or more, about 1.0 mg/mL or more, about It may have an aqueous solubility (or solubility in the coating solution) of 2 mg/mL or more. Combinations of the above-mentioned ranges are possible (e.g., a solubility in water of at least about 10 pg/mL and up to about 1 mg/mL or solubility in the coating solution). Other ranges are also possible. Solid materials may have these or other ranges of water solubility at any point throughout the pH range (eg, pH 1 to pH 14).

In some embodiments, the core may be formed of a material within one of the ranges of solubility classified by the U.S. Pharmacopeia Convention: for example, very soluble:> 1,000 mg/mL; Freely soluble: 100-1,000 mg/mL; Soluble: 33-100 mg/mL; Sparing soluble: 10-33 mg/mL; Slightly soluble: 1-10 mg/mL; Very slightly soluble: 0.1-1 mg/mL; And practically insoluble: <0.1 mg/mL.

Although the core is hydrophobic or hydrophilic, in many embodiments described herein, the core is substantially hydrophobic. “Hydrophilic” and “hydrophilic” are given their ordinary meaning in the art and are, in many cases, relative terms herein, as understood by one of ordinary skill in the art. The relative hydrophobicity and hydrophilicity of a material can be determined by measuring the contact angle of a water droplet on the plane of the material being measured, for example using an instrument such as a contact goniometer and a packed powder of the core material.

In some embodiments, the material (e.g., the material forming the particle core) is at least about 20 degrees, at least about 30 degrees, at least about 40 degrees, at least about 50 degrees, at least about 60 degrees, at least about 70 degrees, about It has a contact angle of 80 degrees or more, about 90 degrees or more, about 100 degrees or more, about 110 degrees or more, about 120 degrees or more, or about 130 degrees or more. In some embodiments, the material is about 160 degrees or less, about 150 degrees or less, about 140 degrees or less, about 130 degrees or less, about 120 degrees or less, about 110 degrees or less, about 100 degrees or less, about 90 degrees or less, about 80 degrees Or less or about 70 degrees or less. Combinations of the above-mentioned ranges are also possible (eg contact angles of about 30 degrees or more and about 120 degrees or less). Other ranges are also possible.

Contact angle measurements can be made using a variety of techniques; Here, the measurement of the static contact angle between the pellets of the starting material and the beads of water that will be used to form the core is mentioned. The material used to form the core was provided as a fine powder or otherwise crushed into fine powder using a mortar and mortar. In order to form a surface to measure against it, the powder was packed using a 7 mm pellet die from International Crystal Labs. The material was added to the die and pressure was applied by hand to pack the powder into the pellets, no pellet press or high pressure was used. The pellets were then suspended for testing so that the top and bottom of the pellets (surface water added and defined as each of the opposite parallel surfaces) did not contact any surface. This was done by not completely removing the pellets from the collar of the die set. Thus, the pellet touches the collar on the side and no contact on the top or bottom. For the contact angle measurement, water was added to the surface of the pellet until a bead of water with a steady contact angle was obtained over 30 seconds. Water was added to the beads of water by immersing or contacting the tip of a pipette or syringe used for the addition of water to the beads. Once stable water beads were obtained, photographs were taken and the contact angle was measured according to standard practice.

In embodiments in which the core comprises an inorganic material (e.g., for use as an imaging agent), the inorganic material is, for example, a metal (e.g., Ag, Au, Pt, Fe, Cr, Co, Ni, Cu, Zn, and other transition metals), semiconductors (e.g. silicon, silicon compounds and alloys, cadmium selenide, cadmium sulfide, indium arsenide, and indium phosphide), or insulators (e.g. ceramics , For example silicon oxide). The inorganic material may be in any suitable amount, for example, at least about 1 wt%, at least about 5 wt%, at least about 10 wt%, at least about 20 wt%, at least about 30 wt%, at least about 40 wt%, about 50 wt. % Or more, about 75 wt% or more, about 90 wt% or more, or about 99 wt% or more in the core. In one embodiment, the core is formed from 100 wt% inorganic material. In some cases, the inorganic material is about 100 wt% or less, about 90 wt% or less, about 80 wt% or less, about 70 wt% or less, about 60 wt% or less, about 50 wt% or less, about 40 wt% or less, about It may be present in the core in an amount of 30 wt% or less, about 20 wt% or less, about 10 wt% or less, about 5 wt% or less, about 2 wt% or less, or about 1 wt% or less. Combinations of the aforementioned ranges are also possible (eg present in an amount of about 1 wt% or more and about 20 wt% or less). Other ranges are also possible.

The core may, in some cases, be in the form of quantum dots, carbon nanotubes, carbon nanowires, or carbon nanorods. In some cases, the core comprises or is formed of a material that is not of biological origin.

In some embodiments, the core comprises one or more organic materials such as synthetic polymers and/or natural polymers. Examples of synthetic polymers include non-degradable polymers such as polymethacrylate and degradable polymers such as polylactic acid, polyglycolic acid and copolymers thereof. Examples of natural polymers include hyaluronic acid, chitosan, and collagen. Other examples of polymers that may be suitable for a portion of the core include those herein suitable for forming coatings on particles, as described below. In some cases, the one or more polymers present in the core can be used to encapsulate or adsorb one or more agents.

In certain embodiments, the core may comprise a medicament comprising lipids and/or proteins. Other materials are also possible.

When the polymer is present in the core, the polymer may be in any suitable amount, for example, about 100 wt% or less, about 90 wt% or less, about 80 wt% or less, about 70 wt% or less, about 60 wt% or less, about 50 wt% % Or less, about 40 wt% or less, about 30 wt% or less, about 20 wt% or less, about 10 wt% or less, about 5 wt% or less, about 2 wt% or less, or about 1 wt% or less in the core . In some cases, the polymer is at least about 1 wt%, at least about 5 wt%, at least about 10 wt%, at least about 20 wt%, at least about 30 wt%, at least about 40 wt%, at least about 50 wt%, about 75 It may be present in the core in an amount of at least about 100 wt%, at least about 90 wt%, or at least about 99 wt%. Combinations of the aforementioned ranges are also possible (eg present in an amount of about 1 wt% or more and about 20 wt% or less). Other ranges are also possible. In one set of embodiments, the core is formed substantially free of polymeric components.

The core can have any suitable shape and/or size. In some cases, the core may be substantially spherical, non-spherical, elliptical, rod-shaped, pyramidal, cube-like, disk-shaped, wire-like, or irregularly shaped. The core is, for example, about 10 μm or less, about 5 μm or less, about 1 μm or less, about 800 nm or less, about 700 nm or less, about 500 nm or less, 400 nm or less, 300 nm or less, about 200 nm or less, about Up to 100 nm or less, about 75 nm or less, about 50 nm or less, about 40 nm or less, about 35 nm or less, about 30 nm or less, about 25 nm or less, about 20 nm or less, about 15 nm or less, or about 5 nm or less Or may have a cross-sectional dimension. In some cases, the core may be, for example, at least about 5 nm, at least about 20 nm, at least about 50 nm, at least about 100 nm, at least about 200 nm, at least about 300 nm, at least about 400 nm, at least about 500 nm, It may have a maximum or minimum cross-sectional dimension of about 1 μm or more or about 5 μm or more. Combinations of the aforementioned ranges are also possible (e.g., a maximum or minimum cross-sectional dimension of at least about 50 nm and less than about 500 nm). Other ranges are also possible. In some embodiments, the size of the core formed by the methods described herein has a Gaussian-type distribution. Unless otherwise specified, measurement of particle/core size herein refers to the minimum cross-sectional dimension.

Those of skill in the art are familiar with techniques for measuring the size of a particle (eg, minimum or maximum cross-sectional dimension). Examples of suitable techniques include (DLS), transmission electron microscopy, scanning electron microscopy, electrical resistance counting, and laser diffraction. Other suitable techniques are known to the skilled person or skilled in the art. Although many methods of determining the size of a particle are known, the sizes (eg, average particle size, thickness) described herein refer to those measured by dynamic light scattering.

Core particle and coated particle formation method

The core particles described herein can be formed by any suitable method. Suitable methods are, for example, so-called top-down techniques, i.e. techniques based on the size reduction of relatively large particles to smaller particles (e.g. milling or homogenization) or bottom-up techniques. , I.e. techniques based on the growth of particles from smaller particles or from individual particles (eg, precipitation or spray-freezing into a liquid).

In some embodiments, the core particles may be coated with a coating. For example, core particles may be provided or formed in a first step, and then the particles may be coated in a second step to form coated particles. In other embodiments, core particles can be formed and coated substantially simultaneously (eg, in a single step). Examples of these and other methods are provided below.

In some embodiments, the coated particles described herein are formed by a method comprising using a formulation process, a milling process, and/or a dilution process. In certain embodiments, the particle formation method comprises a milling process, optionally with a formulation process and/or a dilution process. The formulation process is used to use the core material, one or more surface modifiers, and other ingredients such as solvents, tonicity agents, chelating agents, salts, antimicrobial agents and/or buffers (e.g., sodium citrate and citric acid buffers) (each of which May form a suspension or solution comprising: as described herein. The formulation process can be carried out using a formulation vessel. The core material and other ingredients can be added to the formulation container simultaneously or at a staggered time. The mixture of the core material and/or one or more other ingredients is stirred and/or agitated, or otherwise agitated, in a container to facilitate suspending and/or dissolving the ingredients. In addition, the temperature and/or pressure of the fluid containing the core material, other components, and/or mixtures may be individually increased or decreased to facilitate the suspension and/or dissolution process. In some embodiments, the core material and other components are processed as described herein under an inert atmosphere (eg, nitrogen or argon) and/or in a light-blocked formulation container. The suspension or solution obtained from the formulation container can subsequently be subjected to a milling process followed by a dilution process.

In some embodiments involving a core comprising a solid material, a milling process can be used to reduce the size of the solid material to form particles in the micrometer to nanometer particle size range. The milling process can be carried out using a mill or other suitable instrument. Dry and wet milling processes, such as jet milling, cryo-milling, ball milling, sonication, media milling, and homogenization, are known and are by the methods described herein. Can be used. Generally, in a wet milling process, a suspension of the material used as the core is stirred and mixed with or without excipients to reduce the particle size. Dry milling is a process in which a material used as a core is mixed with a milling medium with or without excipients to reduce particle size. In the cryo-milling process, a suspension of the material used as the core is mixed with the milling medium with or without excipients under cooled temperature.

After milling or other suitable process to reduce the size of the core material, a dilution process can be used to form and/or modify the coated particles from the suspension. The coated particles may comprise a core material, one or more surface modifiers, and other ingredients such as solvents, tonicity agents, chelating agents, salts, antimicrobial agents, and buffering agents (e.g., sodium citrate and citric acid buffers). . By diluting the solution or suspension of the coated particles during the milling step, with or without the addition of surface modifiers and/or other ingredients, the dilution process can be used to achieve the target dosage concentration. In certain embodiments, a dilution process can be used to replace the first surface modifier from the surface of the particle with the second surface modifier as described herein.

The dilution process can be carried out using a product container or any other suitable apparatus. In certain embodiments, the suspension is diluted, i.e., mixed with a diluent or otherwise processed, in a product container. Diluents may contain solvents, surface modifiers, tonicity agents, chelating agents, salts, or antimicrobial agents, or combinations thereof, as described herein. The suspension and diluent can be added to the formulation container simultaneously or at a staggered time. If in certain embodiments the suspension is obtained from a milling process comprising a milling medium, the milling medium can be separated from the suspension prior to adding the suspension to the product container. The suspension, diluent, or mixture of suspension and diluent may be stirred and/or shaken, or otherwise stirred, to form the coated particles described herein. In addition, the temperature and/or pressure of the suspension, diluent, or mixture may be individually increased or decreased to form coated particles. In some embodiments, the suspension and diluent are processed under an inert atmosphere (e.g., nitrogen or argon) and/or in a light-shielded product container.

In some embodiments, the core particles described herein can be prepared by milling a solid material (eg, a medicament) in the presence of one or more stabilizers/surface modifiers. Small particles of solid material require the presence of one or more stabilizers/surface modifiers, especially on the surface of the particles, to stabilize the suspension of particles without agglomeration or aggregation in a liquid solution. can do. In some such embodiments, the stabilizing agent can act as a surface modifier, forming a coating on the particles.

As described herein, in some embodiments, the method of forming core particles includes selecting a stabilizer/surface modifier suitable for milling and also to form a coating on the particles to render the particles mucus penetrating. For example, as described in more detail below, the same rate as the well-established polymer-based MPP due to the 200-500 nm nanoparticles of the model compound pyrene prepared by milling pyrene in the presence of a specific PVA polymer. It was demonstrated that it resulted in particles capable of penetrating the physiological mucus sample. Interestingly, it was observed that only a subset of the PVA polymers tested met the criteria suitable for forming a coating on the particles, also for milling and for mucus penetrating the particles, as described in more detail below.

In a wet milling process, a dispersion (e.g., a dispersion containing one or more stabilizers (e.g., surface modifiers), a grinding medium, a solid to be milled (e.g., a solid drug), and a solvent (e.g., Milling can be performed in an aqueous dispersion). Any suitable amount of stabilizer/surface modifier may be included in the solvent. In some embodiments, the stabilizer/surface modifier is at least about 0.001% (wt% or% weight by volume (w:v)) of the solvent, at least about 0.01%, at least about 0.1%, at least about 0.5%, about 1% or more, about 2% or more, about 3% or more, about 4% or more, about 5% or more, about 6% or more, about 7% or more, about 8% or more, about 10% or more, about 12% or more, about It may be present in the solvent in an amount of at least 15%, at least about 20%, at least about 40%, at least about 60%, or at least about 80%. In some cases, the stabilizer/surface modifier may be present in the solvent in an amount of about 100% (eg, if the stabilizer/surface modifier is a solvent). In other embodiments, the stabilizer/surface modifier is about 100% or less, about 80% or less, about 60% or less, about 40% or less, about 20% or less, about 15% or less, about 12% or less of the solvent, about 10% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less. have. Combinations of the above-mentioned ranges are also possible (eg, an amount of about 5% or less of about 1% or more of the solvent). Other ranges are also possible. The specific range selected is a factor that can affect the ability of the particles to penetrate mucus, such as the stability of the coating of the stabilizer/surface modifier on the particle surface, the average thickness of the coating of the stabilizer/surface modifier on the particles, the particles. The orientation of the stabilizer/surface modifier of the phase, the density of the stabilizer/surface modifier on the particles, the stabilizer:drug ratio, drug concentration, the size and polydispersity of the formed particles, and the morphology of the formed particles can be affected.

The agent (or salt thereof) may be present in the solvent in any suitable amount. In some embodiments, the medicament (or salt thereof) is at least about 0.001% (wt% or% weight by volume (w:v)) of the solvent, at least about 0.01%, at least about 0.1%, at least about 0.5%, about 1% or more, about 2% or more, about 3% or more, about 4% or more, about 5% or more, about 6% or more, about 7% or more, about 8% or more, about 10% or more, about 12% or more, about It is present in the solvent in an amount of at least 15%, at least about 20%, at least about 40%, at least about 60%, or at least about 80%. In some cases, the agent (or salt thereof) is about 100% or less, about 90% or less, about 80% or less, about 60% or less, about 40% or less, about 20% or less, about 15% or less, about 12 or less of the solvent. % Or less, about 10% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less May be present in a solvent. Combinations of the aforementioned ranges are also possible (e.g., an amount of about 20% or less of about 1% or more of the solvent). In some embodiments, the agent is present in the above range but w:v.

The ratio of the stabilizer/surface modifier to the agent (or salt thereof) in the solvent can also be varied. In some embodiments, the ratio of stabilizer/surface modifier to drug (or salt thereof) is at least 0.001:1 (weight ratio, molar ratio, or w:v ratio), at least 0.01:1, at least 0.01:1, 1: It may be 1 or more, 2:1 or more, 3:1 or more, 5:1 or more, 10:1 or more, 25:1 or more, 50:1 or more, 100:1 or more, or 500:1 or more. In some cases, the ratio of stabilizer/surface modifier to drug (or salt thereof) is 1000:1 or less (weight ratio or molar ratio), 500:1 or less, 100:1 or less, 75:1 or less, 50:1 or less , 25:1 or less, 10:1 or less, 5:1 or less, 3:1 or less, 2:1 or less, 1:1 or less, or 0.1:1 or less. Combinations of the above-mentioned ranges are possible (eg, 5:1 or more and 50:1 or less). Other ranges are also possible.

Stabilizers/surface modifiers can be, for example, polymers or surfactants. Examples of polymers are those suitable for use in coatings as described in more detail below. Non-limiting examples of surfactants suitable for use in coatings as surface modifiers include L-α-phosphatidylcholine (PC), 1,2-dipalmitoylphosphatidylcholine (DPPC), oleic acid, sorbitan trioleate, sorbitan mono. Oleate, sorbitan monolaurate, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, natural lecithin, oleyl polyoxyethylene ether, stearyl polyoxyethylene ether, lauryl polyoxyethylene ether , Block copolymer of oxyethylene and oxypropylene, synthetic lecithin, diethylene glycol dioleate, tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl monooleate, glyceryl monostearate, glyceryl Ryl monoricinolate, cetyl alcohol, stearyl alcohol, polyethylene glycol 400, cetyl pyridinium chloride, benzalkonium chloride, olive oil, glyceryl monolaurate, corn oil, cottonseed oil, and sunflower seed oil. Derivatives of the above-mentioned compounds are also possible. In addition, combinations of the above-mentioned compounds and others described herein can be used as surface modifiers in the particles of the present invention. As described herein, in some embodiments the surface modifier may act as a stabilizer, surfactant, and/or emulsifier. In some embodiments, the surface modifier can aid in particle transport in mucus.

In some embodiments, the stabilizer/surface modifier used for milling forms a coating on the particle surface, which coating makes the particles mucus penetrating, while in other embodiments, the stabilizer/surface modifier It should be noted that after is formed, it can be replaced with one or more other stabilizers/surface modifiers. For example, in one set of methods, a first stabilizer/surface modifier may be used during the milling process and may coat the surface of the core particles, followed by removing all or part of the first stabilizer/surface modifier. 2 It is possible to coat all or part of the surface of the core particle by replacing it with a stabilizer/surface modifier. In some cases, the second stabilizer/surface modifier may render the particles more mucus penetrating than the first stabilizer/surface modifier. In some embodiments, it is possible to form core particles with coatings comprising multiple surface modifiers.

Any suitable grinding media can be used for milling. In some embodiments, ceramic and/or polymeric materials and/or metals may be used. Examples of suitable materials may include zirconium oxide, silicon carbide, silicon oxide, silicon nitride, zirconium silicate, yttrium oxide, glass, alumina, alpha-alumina, aluminum oxide, polystyrene, poly(methyl methacrylate), titanium, steel. have. The grinding media can have any suitable size. For example, the grinding media may have an average diameter of at least about 0.1 mm, at least about 0.2 mm, at least about 0.5 mm, at least about 0.8 mm, at least about 1 mm, at least about 2 mm, or at least about 5 mm. In some cases, the grinding media may have an average diameter of about 5 mm or less, about 2 mm or less, about 1 mm or less, about 0.8 mm or less, about 0.5 mm or less, or about 0.2 mm or less. Combinations of the aforementioned ranges are also possible (e.g., an average diameter of about 0.5 millimeters or more and about 1 mm or less). Other ranges are also possible.

Any suitable solvent can be used for milling. The choice of solvent will depend, among other factors, the solid material being milled (e.g., pharmaceutical), the particular type of stabilizer/surface modifier used (e.g., what can make the particles mucus penetrating), It can be determined by factors such as the grinding material used. Suitable solvents may be those that do not substantially dissolve the solid material or the ground material, but dissolve the stabilizer/surface modifier to a suitable degree. Non-limiting examples of solvents include other ingredients such as pharmaceutical excipients, polymers, medicaments, salts, preservatives, viscosity modifiers, tonicity modifiers, taste masking agents, antioxidants, pH modifiers, and Water, buffered solutions, other aqueous solutions, alcohols (eg, ethanol, methanol, butanol), and mixtures thereof, which may optionally contain other pharmaceutical excipients. In other embodiments, organic solvents can be used. The medicament may have any suitable solubility in these or other solvents, such as for solubility in water or in one or more of the ranges described above for solubility in the coating solution.

In other embodiments, the core particles can be formed by an emulsification technique (emulsification). In general, the emulsification technique may involve dissolving or dispersing the material used as the core in a solvent; The solution or dispersion is then emulsified in a second immiscible solvent, thereby forming a number of particles comprising the substance. Suitable emulsification techniques are, for example, by evaporation or extraction, with or without subsequent solvent removal, oil-in-water emulsions, water-in-oil emulsions, water-oil-water emulsions, oil-water-oil emulsions. , Water-in-oil-in-solid emulsions, and oil-in-water-in-solid emulsions, and the like. The emulsification technique is general purpose and can be useful for preparing core particles comprising not only drugs having relatively low water solubility, but also drugs having relatively high water solubility.

In some embodiments, the core particles described herein can be prepared by emulsification in the presence of one or more stabilizers. In some such embodiments, the stabilizer can act as a surface modifier to form a coating on the particles (ie, the emulsifying and coating steps can be performed substantially simultaneously).

In some embodiments, the method of forming core particles by emulsification includes selecting a stabilizer suitable for emulsification and also to form a coating on the particles and render the particles mucus penetrating. For example, as described in more detail below, a physiological mucus sample was prepared at the same rate as the polymer-based MPP settled due to the 200-500 nm nanoparticles of the model polymer PLA prepared by emulsification in the presence of a specific PVA polymer. It has been demonstrated that penetrating particles resulted. Interestingly, it was observed that only a subset of the PVA polymers tested met criteria suitable for emulsification and also for forming coatings on particles, which render the particles mucus penetrating, as described in more detail below.

In another embodiment, the particles are first formed using an emulsification technique, and then the particles are coated with a surface modifier.

Any suitable solvent and solvent combination can be used for the emulsification. Some examples of solvents that can serve as the oil phase include organic solvents such as chloroform, dichloromethane, ethyl acetate, ethyl ether, petroleum ether (hexane, heptane), and oils such as peanut oil, cottonseed oil; Safflower oil; Sesame oil; olive oil; Corn oil, soybean oil, and silicone oil. Some examples of solvents that can serve as the aqueous phase are water and aqueous buffers. Other solvents are also possible.

In other embodiments, core particles can be formed by precipitation techniques. Precipitation techniques (e.g., microprecipitation techniques, nanoprecipitation techniques, crystallization techniques, controlled crystallization techniques) involve forming a first solution comprising a material (e.g., a drug) and a solvent used as a core, , Where the substance is substantially soluble in a solvent. The solution can be added to a second solution comprising another solvent, wherein the material is substantially insoluble (ie, anti-solvent), thereby forming a number of particles comprising the material. In some cases, one or more surface modifiers, surfactants, substances, and/or bioactive agents may be present in the first and/or second solutions. The coating may be formed during the process of sedimenting the core (eg, the sedimentation and coating steps may be performed substantially simultaneously). In other embodiments, the particles are first formed using a precipitation technique and then the particles are coated with a surface modifier.

In some embodiments, precipitation techniques can be used to form polymeric core particles with or without an agent. In general, the precipitation technique dissolves the polymer used as the core in a solvent (with or without an agent present), and then the solution is added to a miscible anti-solvent (with or without an excipient present) to form the core particles. Includes telling. In some embodiments, such techniques are difficult to dissolve (1-10 mg/L), very difficult to dissolve (0.1-1 mg/mL) or in aqueous solutions (e.g., agents with relatively low water solubility), for example It may be useful to prepare polymeric core particles comprising a sparingly insoluble (<0.1 mg/mL) drug.

Any suitable solvent can be used for precipitation. In some embodiments, suitable solvents for precipitation may include, for example, acetone, acetonitrile, dimethylformamide, dimethylsulfoxide, N-methyl-2-pyrrolidone, 2-pyrrolidone, tetrahydrofuran. have. Other organic and non-organic solvents can also be used.

Any suitable anti-solvent can be used for precipitation, including the solvents described herein that can be used for milling. In one set of embodiments, an aqueous solution (e.g., water, buffer solution, other aqueous solution, alcohol (e.g., ethanol, methanol, butanol), which may optionally contain other ingredients such as pharmaceutical excipients, polymers, and medicaments. ), and mixtures thereof).

Stabilizers/surface modifiers for emulsification and precipitation may be polymers or surfactants, including stabilizers/surface modifiers described herein that may be used in milling.

Non-limiting examples of suitable polymers suitable for forming all or part of the core by emulsification or precipitation include polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, polystyrenes, polyimides, Polysulfone, polyurethane, polyacetylene, polyethylene, polyethyleneimine, polyisocyanate, polyacrylate, polymethacrylate, polyacrylonitrile, polyarylate, polypeptide, polynucleotide, and polysaccharide. Non-limiting examples of specific polymers are poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA) , Poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) ( PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO -Co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacrylate, polyurethane, poly-L-lysine ( PLL), hydroxypropyl methacrylate (HPMA), poly(ethylene glycol), poly-L-glutamic acid, poly(hydroxy acid), polyanhydride, polyorthoester, poly(ester amide), polyamide, poly( Ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as Poly(ethylene terephthalate), polyvinyl alcohol (PVA), polyvinyl ether, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxane , Polystyrene (PS), polyurethane, derivatized cellulose, such as alkyl cellulose, hydroxyalkyl cellulose, cellulose ether, cellulose ester, nitrocellulose, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acid, such as poly(methyl(meth )Acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly (Isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly (Isobutyl acrylate), poly(octadecyl acrylate) (collectively referred to herein as "polyacrylic acid"), and copolymers and mixtures thereof, polydioxane and copolymers thereof, polyhydroxyalkanoate, Polypropylene fumarate), polyoxymethylene, poloxamer, poly(ortho)ester, poly(butyric acid), poly(valic acid), poly(lactide-co-caprolactone), and trimethylene carbonate, polyvinyl It includes pyrrolidone, bovine serum albumin, human serum albumin, collagen, DNA, RNA, carboxymethyl cellulose, chitosan, and dextrin.

In some embodiments, precipitation techniques can be used to form particles (e.g., nanocrystals) composed primarily of a drug. In general, such precipitation techniques involve dissolving the polymer used as the core in a solvent and then adding it to a miscible anti-solvent with or without excipients to form core particles. In some embodiments, such techniques are difficult to dissolve (1-10 mg/L), very difficult to dissolve (0.1-1 mg/mL) or in aqueous solutions (e.g., agents with relatively low water solubility), for example It can be useful for preparing particles of a drug that is hardly soluble (<0.1 mg/mL).

In some embodiments, precipitation by salt (or complex) formation can be used to form particles (eg, nanocrystals) of a salt of a drug. In general, precipitation by salt formation dissolves a substance used as a core in a solvent with or without excipients, followed by addition of counter-ions or complexing agents forming an insoluble salt or complex of the drug to form core particles. Includes that. This technique can be useful for preparing particles of a medicament soluble in aqueous solution (eg, an agent with a relatively high water solubility). In some embodiments, agents having one or more charged or ionizable groups can interact with counter-ions (eg, cations or anions) to form salt complexes.

A variety of counter-ions can be used to form salt complexes, including metals (eg, alkali metals, alkaline earth metals and transition metals). Non-limiting examples of cationic counter-ions include zinc, calcium, aluminum, zinc, barium, and aluminum. Non-limiting examples of anionic counter-ions include phosphates, carbonates, and fatty acids. Counter-ions can be monovalent, divalent, or trivalent, for example. Other counter-ions are known in the art and can be used in the embodiments described herein. Other ionic and nonionic complexing agents are also possible.

A variety of different acids can be used in the precipitation process. In some embodiments, suitable acids may include decanoic acid, hexanoic acid, mucinic acid, octanoic acid. In other embodiments, suitable acids are acetic acid, adipic acid, L-ascorbic acid, L-aspartic acid, capric acid (decanoic acid), carbonic acid, citric acid, fumaric acid, galactaric acid, D-glucoheptonic acid, D-glu Conic acid, D-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrochloric acid, DL-lactic acid, lauric acid, maleic acid, (-)-L-malic acid, palmitic acid Acid, phosphoric acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, (+)-L-tartaric acid, or thiocyanic acid. In other embodiments, suitable acids are alginic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, caprylic acid (octanoic acid), cyclamic acid, dodecylsulfonic acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, Ethanesulfonic acid, 2-hydroxy-, gentisic acid, glutaric acid, 2-oxo-, isobutyric acid, lactobionic acid, malonic acid, methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 2-naphthoic acid, 1-hydroxy-, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmic acid, (embonic acid), propionic acid, (-)-L-pyroglutamic acid, or p -toluenesulfonic acid. In another embodiment suitable acids are acetic acid, 2,2-dichloro-, benzoic acid, 4-acetamido-, (+)-camphor-10-sulfonic acid, caproic acid (hexanoic acid), cinnamic acid, formic acid, brominated Hydrogen acid, DL-mandelic acid, nitric acid, salicylic acid, salicylic acid, 4-amino-, or undecylenic acid (undes-10-enoic acid). Mixtures of one or more such acids may also be used.

A variety of different bases can be used in the precipitation process. In some embodiments, suitable bases include ammonia, L-arginine, calcium hydroxide, choline, glucamine, N -methyl-, lysine, magnesium hydroxide, potassium hydroxide, or sodium hydroxide. In other embodiments, suitable bases are benetamine, benzathine, betaine, theanol, diethylamine, ethanol, 2-(diethylamino)-, hydravamine, morpholine, 4-(2-hydroxyethyl )-Morpholine, pyrrolidine, 1-(2-hydroxyethyl)-, or tromethamine. In other embodiments, suitable bases are diethanolamine (2,2'-iminobis(ethanol)), ethanolamine (2-aminoethanol), ethylenediamine, 1 H -imidazole, piperazine, triethanolamine (2, 2',2"-nitrilotris (ethanol)), or zinc hydroxide. Mixtures of one or more of these bases may also be used.

Any suitable solvent can be used for precipitation by salt formation, including the solvents described herein that can be used for milling. In one set of embodiments, aqueous solutions (e.g., water, buffered solutions, other aqueous solutions, and alcohols (e.g., ethanol, methanol, Butanol)), and mixtures thereof.

In the precipitation process, the salt may have a lower aqueous solubility (or solubility in the solvent containing the salt) than the non-salt form of the drug. The aqueous solubility (or solubility in a solvent) of the salt is, for example, about 5 mg/mL or less, about 2 mg/mL or less, about 1 mg/mL or less, about 0.5 mg/mL or less, about 0.1 mg/mL at 25°C. Or less, about 0.05 mg/mL, about 0.01 mg/mL or less, about 1 μg/mL or less, about 0.1 μg/mL or less, about 0.01 μg/mL or less, about 1 ng/mL or less, about 0.1 ng/mL or less, or It may be less than or equal to about 0.01 ng/mL. In some embodiments, the salt is at least about 1 pg/mL, at least about 10 pg/mL, at least about 0.1 ng/mL, at least about 1 ng/mL, at least about 10 ng/mL, at least about 0.1 μg/mL, about 1 μg/mL or more, about 5 μg/mL or more, about 0.01 mg/mL or more, about 0.05 mg/mL or more, about 0.1 mg/mL or more, about 0.5 mg/mL or more, about 1.0 mg/mL or more, about 2 It may have an aqueous solubility (or solubility in a solvent) of mg/mL. Combinations of the aforementioned ranges are possible (e.g., an aqueous solubility (or solubility in a solvent) of at least about 0.001 mg/mL and up to about 1 mg/mL). Other ranges are also possible. Salts may have these or other ranges of water solubility at any point throughout the pH range (eg, pH 1 to pH 14).

In some embodiments, the solvent used for precipitation comprises one or more surface modifiers as described herein, and as it precipitates from solution, a coating of one or more surface modifiers may form around the particles. The surface modifier is in any suitable concentration, such as at least about 0.001% (w/v), at least about 0.005% (w/v), at least about 0.01% (w/v), at least about 0.05% (w/v) in aqueous solution. It may be present in the solvent at a concentration of at least about 0.1% (w/v), at least about 0.5% (w/v), at least about 1% (w/v), or at least about 5% (w/v). In some cases, the surface modifier is about 5% (w/v) or less, about 1% (w/v) or less, about 0.5% (w/v) or less, about 0.1% (w/v) or less, about 0.05 % (w/v) or less, about 0.01% (w/v) or less, or about 0.005% (w/v) or less in the solvent. Combinations of the aforementioned ranges are also possible (e.g., concentrations of about 0.01% (w/v) or more and about 1% (w/v) or less). Other ranges are also possible.

Another exemplary method of forming the core particles includes freeze-drying techniques. In this technique, the medicament or its salt can be dissolved in an aqueous solution, optionally containing a surface modifier. Counter-ions can be added to the solution, and the solution can be flash frozen immediately and freeze-dried. The dry powder can be restored in a suitable solvent (eg aqueous solution, such as water) at the desired concentration.

Counter-ions can be added to the solvent for freeze drying in any suitable range. In some cases, the ratio of counter-ion to drug (e.g., salt) is at least 0.1:1 (weight or molar ratio), at least 1:1, at least 2:1, at least 3:1, at least 5:1, It may be 10:1 or more, 25:1 or more, 50:1, or 100:1 or more. In some cases, the ratio of counter-ion to drug (e.g., salt) is 100:1 (weight or molar ratio) or less, 75:1 or less, 50:1 or less, 25:1 or less, 10:1 or less, It may be 5:1 or less, 3:1 or less, 2:1 or less, 1:1 or less, or 0.1:1 or less. Combinations of the above-mentioned ranges are possible (eg ratios of 5:1 to 50:1). Other ranges are also possible.

If the surface modifier is present in the solvent prior to freeze drying, it is at any suitable concentration, such as at least about 0.001% (w/v), at least about 0.005% (w/v), at least about 0.01% (w/v) in aqueous solution. , At least about 0.05% (w/v), at least about 0.1% (w/v), at least about 0.5% (w/v), at least about 1% (w/v), or at least about 5% (w/v) May exist in concentration. In some cases, the surface modifier is about 5% (w/v) or less, about 1% (w/v) or less, about 0.5% (w/v) or less, about 0.1% (w/v) or less, about 0.05 % (w/v) or less, about 0.01% (w/v) or less, or about 0.005% (w/v) or less in the solvent. Combinations of the aforementioned ranges are also possible (e.g., concentrations of about 0.01% (w/v) or more and about 1% (w/v) or less). Other ranges are also possible.

The concentration of the surface-altering agent present in the solvent can be above or below the critical micelle concentration (CMC) of the surface-altering agent, depending on the particular surface-altering agent used. In other embodiments, stable particles can be formed by adding excess counter-ions to a solution containing the drug. The precipitate can then be washed by various methods such as centrifugation. The resulting slurry can be sonicated. One or more surface modifiers may be added to stabilize the resulting particles.

Other methods of forming the core particles are also possible. Core particle formation techniques include, for example, coacervation-phase separation; Melt dispersion; Interfacial deposition; In situ polymerization; Self-assembly of macromolecules (eg, formation of a polyelectrolyte complex or a polyelectrolyte-surfactant complex); Spray-drying and spray-congealing; Electro-spray; Air suspension coating; Pan and spray coating; Freeze-drying, air drying, vacuum drying, fluidized-bed drying; Precipitation (eg, nanoprecipitation, microprecipitation); Critical fluid extraction; And a lithographic approach (eg, soft lithography, step and flash imprint lithography, interference lithography, photolithography).

Combinations of the methods described herein with other methods are also possible. For example, in some embodiments, the core of the medicament is first formed by precipitation and then the size of the core is further reduced by a milling process.

After particle formation of the drug, the particles can be optionally exposed to a solution comprising a (second) surface-altering agent that can associate with and/or coat the particles. In embodiments where the medicament already comprises a coating of the first surface modifying agent, all or part of the second surface modifying agent may be replaced with the second stabilizer/surface modifying agent to coat all or part of the particle surface. In some cases, the second surface modifier may render the particles more mucus penetrating than the first surface modifier. In other embodiments, particles with coatings comprising multiple surface modifiers can be formed (eg, in a single layer or in multiple layers). In other embodiments, particles with multiple coatings can be formed (eg, each coating optionally includes a different surface modifier). In some cases, the coating is in the form of a single layer of surface modifier. Other configurations are also possible.

In any one of the methods described herein, at least about 1 minute, at least about 2 minutes, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 30 minutes, at least about 60 minutes, Alternatively, the particles can be coated with a surface-altering agent by incubating the particles in solution with a surface-altering agent for an extended period of time. In some cases, the incubation may be conducted for a period of about 10 hours or less, about 5 hours or less, or about 60 minutes or less. Combinations of the aforementioned ranges are also possible (e.g., an incubation period of about 2 minutes or more up to 60 minutes).

Particle coating

As shown in the embodiment illustrated in FIG. 1, the core 16 may be surrounded by a coating 20 comprising one or more surface modifiers. In some embodiments, the coating is formed from one or more surface modifiers or other molecules disposed on the surface of the core. The specific chemical composition and/or component of the coating and surface modifier(s) can be selected to impart specific functionality to the particles, such as improved transport through the mucosal barrier.

It should be understood that the coating surrounding the core need not completely surround the core, but such embodiments may be possible. For example, the coating may be at least about 10%, at least about 30%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 99% of the surface area of the core. Can surround In some cases, the coating substantially surrounds the core. In other cases, the coating completely surrounds the core. In other embodiments, the coating covers about 100% or less, about 90% or less, about 80% or less, about 70% or less, about 60% or less, or about 50% or less of the surface area of the core. Combinations of the above mentioned ranges are also possible (eg, covering more than 80% and less than 100% of the surface area of the core).

The components of the coating may be evenly distributed over the surface of the core in some cases and unevenly in other cases. For example, the coating may in some cases include portions that do not contain any material (eg, hollow). If desired, the coating can be designed to allow certain molecules and components to penetrate and/or transport into or out of the coating, but prevent other molecules and components from penetrating and/or transporting into or out of the coating. have. The ability of a particular molecule to penetrate and/or be transported into and/or across the coating depends, for example, on the packing density of the surface modifier forming the coating and the chemical and chemical properties of the components forming the coating. May vary according to physical properties. As described herein, the coating may, in some embodiments, comprise one layer of material (eg, a single layer), or multiple layers of material. There may be a single type of surface modifying agent, or multiple types of surface modifying agents.

The coating of particles can have any suitable thickness. For example, the coating is about 1 nm or more, about 5 nm or more, about 10 nm or more, about 30 nm or more, about 50 nm or more, about 100 nm or more, about 200 nm or more, about 500 nm or more, about 1 μm It may have an average thickness of greater than or equal to about 5 μm. In some cases, the average thickness of the coating is about 5 μm or less, about 1 μm or less, about 500 nm or less, about 200 nm or less, about 100 nm or less, about 50 nm or less, about 30 nm or less, about 10 nm or less, or about Less than 5 nm. Combinations of the above-mentioned ranges are also possible (eg an average thickness of about 1 nm or more and about 100 nm or less). Other ranges are also possible. In the case of particles with multiple coatings, each coating layer may have one of the thicknesses described above.

In some embodiments, the compositions and methods described herein may make it possible to coat core particles with a hydrophilic surface modifier without the need for covalent linking of surface modifying moieties to the core surface. In some such embodiments, a core having a hydrophobic surface may be coated with a polymer described herein, resulting in a number of surface modifying moieties on the core surface without substantially altering the characteristics of the core itself. For example, a surface modifier can be adsorbed to the outer surface of the core particles. However, in other embodiments, the surface modifier is covalently linked to the core particle. In certain embodiments in which the surface modifier is adsorbed onto the surface of the core, the surface modifier may, optionally, equilibrate with other components (eg, in the composition/formulation) and other molecules of the surface modifier in solution. In some cases, the adsorbed surface modifier may be present on the surface of the core at the densities described herein. The density can be an average density because the surface modifier is in equilibrium with the other components in the solution.

The coatings and/or surface modifiers of the particles described herein may comprise any suitable material, such as a hydrophobic material, a hydrophilic material, and/or an amphiphilic material. In some embodiments, the coating comprises a polymer. In certain embodiments, the polymer is a synthetic polymer (ie, a polymer that is not naturally produced). In other embodiments, the polymer is a natural polymer (eg, protein, polysaccharide, rubber). In certain embodiments, the polymer is a surface active polymer. In certain embodiments, the polymer is a nonionic polymer. In certain embodiments, the polymer is a linear, synthetic nonionic polymer. In certain embodiments, the polymer is a nonionic block copolymer. In some embodiments, the polymer can be, for example, a copolymer in which one repeat unit is relatively hydrophobic and another repeat unit is relatively hydrophilic. The copolymer may be, for example, a diblock, triblock, alternating, or random copolymer. The polymer may or may not be charged.

In some embodiments, the coating comprises a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer. For example, in certain embodiments, the polymer may comprise poly(vinyl alcohol), partially hydrolyzed poly(vinyl acetate), or a copolymer of vinyl alcohol and vinyl acetate. In certain embodiments, the synthetic polymer having pendant hydroxyl groups on the backbone of the polymer is a poly(ethylene glycol)-poly(vinyl acetate)-poly(vinyl alcohol) copolymer, a poly(ethylene glycol)-poly(vinyl alcohol) copolymer. Coalescence, poly(propylene oxide)-poly(vinyl alcohol) copolymer, and poly(vinyl alcohol)-poly(acryl amide) copolymer. Without wishing to be bound by theory, particles comprising a coating comprising a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer, at least in part, due to the display of a number of hydroxyl groups on the particle surface, It may have reduced mucosal adhesion compared to the control particles. One possible mechanism for reduced mucoadhesion is that the hydroxyl groups alter the microenvironment of the particles, for example by ordering water and other molecules in the particle/mucous environment. An additional or alternative possible mechanism is that the hydroxyl groups mask the adherent domain of the mucin fibers, thereby reducing particle adhesion and speeding up particle transport.

Moreover, the ability of particles coated with synthetic polymers having pendant hydroxyl groups on the backbone of the polymer to penetrate mucus can also be determined, at least in part, by the degree of hydrolysis of the polymer. In some embodiments, the hydrophobic portion of the polymer (e.g., the portion of the polymer that does not hydrolyze) can make the polymer attachable to the core surface (e.g., if the core surface is hydrophobic), and thus the core It enables a strong association between the polymer and the polymer. Surprisingly, in some embodiments involving the surface modifier PVA, too high a degree of hydrolysis makes sufficient adhesion between the PVA and the core not possible (e.g., if the core is hydrophobic), and thus coating with such a polymer. It has been found that the resulting particles generally do not exhibit sufficiently reduced mucosal adhesion. In some embodiments, too low a degree of hydrolysis does not enhance particle transport in mucus, possibly due to the lower amount of hydroxyl groups available to alter the microenvironment of the particles and/or mask the adherent domains of mucin fibers. .

Synthetic polymers having pendant hydroxyl groups on the backbone of the polymer may have any suitable degree of polymerization (and thus varying amounts of hydroxyl groups). The appropriate level of hydrolysis can be determined by additional factors such as the molecular weight of the polymer, the composition of the core, the hydrophobicity of the core, and the like. In some embodiments, the synthetic polymer (e.g., PVA or partially hydrolyzed poly(vinyl acetate) or a copolymer of vinyl alcohol and vinyl acetate) is at least about 30% hydrolyzed, at least about 35% hydrolyzed, about More than 40% hydrolysis, more than about 45% hydrolysis, more than about 50% hydrolysis, more than about 55% hydrolysis, more than about 60% hydrolysis, more than about 65% hydrolysis, more than about 70% hydrolysis, about 75 % Or more hydrolysis, about 80% or more hydrolysis, about 85% or more hydrolysis, about 87% or more hydrolysis, about 90% or more hydrolysis, about 95% or more hydrolysis, or about 98% or more hydrolysis . In some embodiments, the synthetic polymer is about 100% or less hydrolyzed, about 98% or less hydrolyzed, about 97% or less hydrolyzed, about 96% or less hydrolyzed, about 95% or less hydrolyzed, about 94% or less hydrolyzed, Less than about 93% hydrolysis, less than about 92% hydrolysis, less than about 91% hydrolysis, less than about 90% hydrolysis, less than about 87% hydrolysis, less than about 85% hydrolysis, less than about 80% hydrolysis, about Less than 75% hydrolysis, less than about 70% hydrolysis, or less than about 60% hydrolysis. Combinations of the aforementioned ranges are also possible (eg, about 80% or more hydrolyzed and about 95% or less hydrolyzed polymer). Other ranges are also possible.

The molecular weight of the synthetic polymers described herein (e.g., those having pendant hydroxyl groups on the backbone of the polymer) can be selected to reduce mucoadhesion of the core and ensure sufficient association of the polymer with the core. In certain embodiments, the molecular weight of the synthetic polymer is about 1 kDa or more, about 2 kDa or more, about 5 kDa or more, about 8 kDa or more, about 9 kDa about 10 kDa or more, about 12 kDa or more, about 15 kDa or more, about 20 kDa or more, about 25 kDa or more, about 30 kDa or more, about 40 kDa or more, about 50 kDa or more, about 60 kDa or more, about 70 kDa or more, about 80 kDa or more, about 90 kDa or more, about 100 kDa or more, about 110 kDa or more, about 120 kDa or more, about 130 kDa or more, about 140 kDa or more, about 150 kDa or more, about 200 kDa or more, about 500 kDa or more, or about 1000 kDa or more. In some embodiments, the molecular weight of the synthetic polymer is about 1000 kDa or less, about 500 kDa or less, about 200 kDa or less, about 180 kDa or less, about 150 kDa or less, about 130 kDa or less, about 120 kDa or less, about 100 kDa or less, About 85 kDa or less, about 70 kDa or less, about 65 kDa or less, about 60 kDa or less, about 50 kDa or less, or about 40 kDa or less, about 30 kDa or less, about 20 kDa or less, about 15 kDa or less, or about 10 kDa or less . Combinations of the above-mentioned ranges are also possible (eg molecular weights of about 10 kDa or more and about 30 kDa or less). The aforementioned molecular weight range can also be combined with the aforementioned hydrolysis range to form suitable polymers.

In some embodiments, the synthetic polymers described herein are or comprise PVA. PVA is a nonionic polymer with surface active properties. It is a synthetic polymer that is typically prepared through hydrolysis of poly(vinyl acetate). The partially hydrolyzed PVA consists of two types of repeat units, a vinyl alcohol unit and the remaining vinyl acetate units. Vinyl alcohol units are relatively hydrophilic; Vinyl acetate units are relatively hydrophobic. In some cases, the sequence distribution of vinyl alcohol units and vinyl acetate units is blocky. For example, a series of vinyl alcohol units, followed by a series of vinyl acetate units, and then further vinyl alcohol units, can form a polymer having a mixed block-copolymer type arrangement, with units distributed in a blocky manner. have. In certain embodiments, the repeating units form a copolymer, such as a diblock, triblock, alternating, or random copolymer. Polymers other than PVA may also have these configurations of hydrophilic units and hydrophobic units.

In certain embodiments, the surface modifier is a PVA having a molecular weight of about 98% or less and a molecular weight of about 75 kDa or less, or less than about 95% hydrolyzed PVA. In some embodiments, the surface modifier is a PVA that does not have both properties of a degree of hydrolysis greater than 95% and a molecular weight greater than 31 kDa. In certain embodiments, such surface modifiers can be used to coat certain agents, such as corticosteroids (eg, LE) and/or other compounds described herein.

In some embodiments, the hydrophilic units of the synthetic polymers described herein may be present substantially on the outer surface of the particle. For example, hydrophilic units can form most of the outer surface of the coating and can help stabilize the particles in an aqueous solution containing the particles. Hydrophobic units may be present substantially inside the coating and/or at the surface of the core particles, for example to facilitate adhesion of the coating to the core.

The molar fractions of the relatively hydrophilic units and the relatively hydrophobic units of the synthetic polymer can be selected to reduce mucoadhesion of the core and ensure sufficient association of the polymer with the core, respectively. As described herein, the mole fraction of the hydrophobic units of the polymer can be selected to increase the likelihood that proper association of the polymer with the core occurs, thereby causing the polymer to remain attached to the core. The mole fraction of the relatively hydrophilic unit relative to the relatively hydrophobic unit of the synthetic polymer is, for example, 0.5:1 or more, 1:1 or more, 2:1 or more, 3:1 or more, 5:1 or more, 7:1 or more, It may be 10:1 or more, 15:1 or more, 20:1 or more, 25:1 or more, 30:1 or more, 40:1 or more, 50:1 or more, 75:1 or more, or 100:1 or more. In some embodiments, the mole fraction of relatively hydrophilic units relative to relatively hydrophobic units of the synthetic polymer is, for example, 100:1 or less, 75:1 or less, 50:1 or less, 40:1 or less, 30:1 or less, It may be 25:1 or less, 20:1 or less, 15:1 or less, 10:1 or less, 7:1 or less, 5:1 or less, 3:1 or less, 2:1 or less, or 1:1 or less. Combinations of the aforementioned ranges are also possible (e.g., ratios of 1:1 or more and 50:1 or less). Other ranges are also possible.

The molecular weight of the PVA polymer can also be adjusted to increase the effectiveness of the polymer to allow particles to penetrate mucus. Examples of PVA polymers having various molecular weights and degrees of hydrolysis are shown in Table 1.

In certain embodiments, the synthetic polymer is represented by the formula:

In the above formula, n is all integers from 0 to 22730; m is both an integer from 0 to 11630. In certain embodiments, n is all integers from 25 to 20600. In some embodiments, m is all integers from 5 to 1100. In certain embodiments, m is all integers from 0 to 400 or all integers from 1 to 400. Note that n and m each represent the total content of vinyl alcohol and vinyl acetate repeat units in the polymer, rather than the block length.

The value of n can be different. In certain embodiments, n is 5 or more, 10 or more, 20 or more, 30 or more, 50 or more, 100 or more, 200 or more, 300 or more, 500 or more, 800 or more, 1000 or more, 1200 or more, 1500 or more, 1800 or more, 2000 Or more, 2200 or more, 2400 or more, 2600 or more, 3000 or more, 5000 or more, 10000 or more, 15000 or more, 20000 or more, or 25000 or more. In some cases, n is 30000 or less, 25000 or less, 20000 or less, 25000 or less, 20000 or less, 15000 or less, 10000 or less, 5000 or less, 3000 or less, 2800 or less, 2400 or less, 2000 or less, 1800 or less, 1500 or less, 1200 or less , 1000 or less, 800 or less, 500 or less, 300 or less, 200 or less, 100 or less, or 50 or less. Combinations of the aforementioned ranges are also possible (for example, n is 50 or more and 2000 or less). Other ranges are also possible.

Similarly, the value of m can be different. For example, in certain embodiments, m is 5 or more, 10 or more, 20 or more, 30 or more, 50 or more, 70 or more, 100 or more, 150 or more, 200 or more, 250 or more, 300 or more, 350 or more, 400 or more, 500 or more, 800 or more, 1000 or more, 1200 or more, 1500 or more, 1800 or more, 2000 or more, 2200 or more, 2400 or more, 2600 or more, 3000 or more, 5000 or more, 10000 or more, or 15000 or more. In some cases, m is 15000 or less, 10000 or less, 5000 or less, 3000 or less, 2800 or less, 2400 or less, 2000 or less, 1800 or less, 1500 or less, 1200 or less, 1000 or less, 800 or less, 500 or less, 400 or less, 350 or less , 300 or less, 250 or less, 200 or less, 150 or less, 100 or less, 70 or less, 50 or less, 30 or less, 20 or less, or 10 or less. Combinations of the aforementioned ranges are also possible (for example, m is 5 or more and 200 or less). Other ranges are also possible.

In some embodiments, the particles described herein comprise a coating comprising a block copolymer having a relatively hydrophilic block and a relatively hydrophobic block. In some cases, the hydrophilic block may be substantially present on the outer surface of the particle. For example, hydrophilic blocks can form most of the outer surface of the coating and can help stabilize the particles in an aqueous solution containing the particles. The hydrophobic block may be present substantially inside the coating and/or at the surface of the core particles, for example promoting adhesion of the coating to the core. In some cases, the coating comprises a surface modifier comprising a triblock copolymer, wherein the triblock copolymer comprises a hydrophilic block-hydrophobic block-hydrophilic block configuration. Diblock copolymers having a hydrophilic block-hydrophobic block configuration are also possible. Combinations of block copolymers and other polymers suitable for use as coatings are also possible. Non-linear block configurations, such as comb, brush, or star copolymers are also possible. In some embodiments, the relatively hydrophilic block comprises a synthetic polymer (eg, PVA) having pendant hydroxyl groups on the backbone of the polymer.

The molecular weight of the hydrophilic block and the hydrophobic block of the block copolymer can be selected to reduce mucoadhesion of the core and ensure sufficient association of the block copolymer with the core, respectively. The molecular weight of the hydrophobic block of the block copolymer can be selected to increase the likelihood that proper association of the block copolymer with the core occurs, thereby causing the block copolymer to remain attached to the core.

In certain embodiments, the combined molecular weight of (at least one) relatively hydrophobic block or repeat units of the block copolymer is at least about 0.5 kDa, at least about 1 kDa, at least about 2 kDa, at least about 3 kDa, at least about 4 kDa, About 5 kDa or more, about 6 kDa or more, about 10 kDa or more, about 12 kDa or more, about 15 kDa or more, about 20 kDa or more, or about 50 kDa or more, about 60 kDa or more, about 70 kDa or more, about 80 kDa or more, About 90 kDa or more, about 100 kDa or more, about 110 kDa or more, about 120 kDa or more, about 130 kDa or more, about 140 kDa or more, about 150 kDa or more, about 200 kDa or more, about 500 kDa or more, or about 1000 kDa or more . In some embodiments, the combined molecular weight of (one or more) relatively hydrophobic blocks or repeat units is about 1000 kDa or less, about 500 kDa or less, about 200 kDa or less, about 150 kDa or less, about 140 kDa or less, about 130 kDa or less. , About 120 kDa or less, about 110 kDa or less, about 100 kDa or less, about 90 kDa or less, about 80 kDa or less, about 50 kDa or less, about 20 kDa or less, about 15 kDa or less, about 13 kDa or less, about 12 kDa or less , About 10 kDa or less, about 8 kDa or less, or about 6 kDa or less. Combinations of the above-mentioned ranges are also possible (eg, greater than or equal to about 3 kDa and less than or equal to about 15 kDa). Other ranges are also possible.

In some embodiments, the combined (one or more) relatively hydrophilic block or repeat units of the block copolymer is at least about 15 wt%, at least about 20 wt%, at least about 25 wt%, at least about 30 wt% of the block copolymer. , At least about 35 wt%, at least about 40 wt%, at least about 45 wt%, at least about 50 wt%, at least about 55 wt%, at least about 60 wt%, at least about 65 wt%, or at least about 70 wt% do. In some embodiments, the combined (one or more) relatively hydrophilic block or repeat units of the block copolymer is about 90 wt% or less, about 80 wt% or less, about 60 wt% or less, about 50 wt% or less of the block copolymer. Or about 40 wt% or less. Combinations of the above-mentioned ranges are also possible (eg, about 30 wt% or more and about 80 wt% or less). Other ranges are also possible.

In some embodiments, the combined molecular weight of (at least one) relatively hydrophilic block or repeat units of the block copolymer is at least about 0.5 kDa, at least about 1 kDa, at least about 2 kDa, at least about 3 kDa, at least about 4 kDa, About 5 kDa or more, about 6 kDa or more, about 10 kDa or more, about 12 kDa or more, about 15 kDa or more, about 20 kDa or more, or about 50 kDa or more, about 60 kDa or more, about 70 kDa or more, about 80 kDa or more, About 90 kDa or more, about 100 kDa or more, about 110 kDa or more, about 120 kDa or more, about 130 kDa or more, about 140 kDa or more, about 150 kDa or more, about 200 kDa or more, about 500 kDa or more, or about 1000 kDa or more have. In certain embodiments, the combined molecular weight of (one or more) relatively hydrophilic blocks or repeat units is about 1000 kDa or less, about 500 kDa or less, about 200 kDa or less, about 150 kDa or less, about 140 kDa or less, about 130 kDa or less. , About 120 kDa or less, about 110 kDa or less, about 100 kDa or less, about 90 kDa or less, about 80 kDa or less, about 50 kDa or less, about 20 kDa or less, about 15 kDa or less, about 13 kDa or less, about 12 kDa or less , About 10 kDa or less, about 8 kDa or less, about 6 kDa or less, about 5 kDa or less, about 3 kDa or less, about 2 kDa or less, or about 1 kDa or less. Combinations of the aforementioned ranges are also possible (e.g., greater than or equal to about 0.5 kDa and less than or equal to about 3 kDa). Other ranges are also possible. In embodiments in which the two hydrophilic blocks are flanked by the hydrophobic block, the molecular weights of the two hydrophilic blocks can be substantially the same or different.

In certain embodiments, the polymer of the surface modifier comprises a polyether moiety. In certain embodiments, the polymer comprises a polyalkylether moiety. In certain embodiments, the polymer comprises a polyethylene glycol tail. In certain embodiments, the polymer comprises a polypropylene glycol core. In certain embodiments, the polymer comprises polybutylene glycol as a core. In certain embodiments, the polymer comprises polypentylene glycol as a core. In certain embodiments, the polymer comprises polyhexylene glycol as a core. In certain embodiments, the polymer is a diblock copolymer of one of the polymers described herein. In certain embodiments, the polymer is a triblock copolymer of one of the polymers described herein. As disclosed herein, any description of PEG can be replaced with polyethylene oxide (PEO), and any description of PEO can be replaced with PEG. In some embodiments, the diblock or triblock copolymer is a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer as one or more of the blocks (having varying degrees of hydrolysis and varying molecular weights as described herein) (e.g. , PVA). The synthetic polymer block may form the central or end portion of the block copolymer.

In certain embodiments, the polymer is a polyalkyl ether (e.g., polyethylene glycol, polypropylene glycol) and another polymer (e.g., a synthetic polymer having pendant hydroxyl groups on the backbone of the polymer (e.g., PVA) ) Is a triblock copolymer. In certain embodiments, the polymer is a triblock copolymer of a polyalkyl ether and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer of polyethylene glycol and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer of polypropylene glycol and another polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer having one or more units of a polyalkyl ether. In certain embodiments, the polymer is a triblock copolymer of two different polyalkyl ethers. In certain embodiments, the polymer is a triblock copolymer comprising polyethylene glycol units. In certain embodiments, the polymer is a triblock copolymer comprising polypropylene glycol units. In certain embodiments, the polymer is a triblock copolymer of more hydrophobic units flanked by two or more hydrophilic units. In certain embodiments, the hydrophilic units are of the same type of polymer. In some embodiments, the hydrophilic unit comprises a synthetic polymer (eg, PVA) having pendant hydroxyl groups on the backbone of the polymer. In certain embodiments, the polymer comprises polypropylene glycol units flanked by two or more hydrophilic units. In certain embodiments, the polymer comprises two polyethylene glycol units flanked by more hydrophobic units. In certain embodiments, the polymer is a triblock copolymer having polypropylene glycol units flanked by two polyethylene glycol units. The molecular weights of the two blocks flanking the central block may be substantially the same or different.

In certain embodiments, the polymer is of the formula:

In the above formula, n is all integers from 2 to 1140; m is both an integer from 2 to 1730. In certain embodiments, n is all integers from 10 to 170. In certain embodiments, m is all integers from 5 to 70. In certain embodiments, n is at least 2 times m, at least 3 times m, or at least 4 times m.

In certain embodiments, the coating is present on the coating alone or in combination with a synthetic polymer having pendant hydroxyl groups (e.g., PVA) on another polymer, such as the backbone of the polymer, (poly(ethylene glycol)) -(Poly(propylene oxide))-(poly(ethylene glycol)) triblock copolymer (hereinafter, "PEG-PPO-PEG triblock copolymer"). As described herein, PEG blocks may be interchanged with PEO blocks in some embodiments. The molecular weight of the PEG (or PEO) and PPO segments of the PEG-PPO-PEG triblock copolymer can be selected to reduce the adhesion of the particles, as described herein. Without wishing to be bound by theory, particles having a coating comprising a PEG-PPO-PEG triblock copolymer are, at least in part, due to the display of multiple PEG (or PEO) segments on the particle surface, as a control It may have reduced mucoadhesion compared to particles. PPO segments can be attached to the core surface (eg, if the core surface is hydrophobic), thus allowing strong association between the core and the polymer. In some cases, the PEG-PPO-PEG triblock copolymer is associated with the core through non-covalent interactions. For comparison purposes, the control particles may be, for example, carboxylate-modified polystyrene particles of similar size to the corresponding coated particles.

In certain embodiments, the surface modification agent comprises a polymer containing a poloxamer having a nick ^® under the trade name flu. A fluorenyl, which may be useful in embodiments described herein Nick ^® polymers include F127, F38, F108, F68, F77, F87, F88, F98, L101, L121, L31, L35, L43, L44, L61, L62, L64, L81, L92, N3, P103, P104, P105, P123, P65, P84, and P85 are included, but are not limited thereto.

Examples of the molecular weights of specific Pluronic ^® molecules are shown in Table 2.

Although other ranges are possible and may be useful in certain embodiments described herein, in some embodiments, the hydrophobic block of the PEG-PPO-PEG triblock copolymer has one of the molecular weights described above (e.g., about 3 kDa or more and about 15 kDa or less), the combined hydrophilic block has a weight percentage relative to the polymer in one of the ranges described above (e.g., about 15 wt% or more, about 20 wt% or more, about 25 wt% or more, or about At least 30 wt%, and up to about 80 wt%). Nick ^® polymer to a specific fluorene belonging to these standards include, for example, the F127, F108, P105, and P103.

The ability of the surface-modifying agent to make the particle or core mucus penetrating is at least in part, the ability of the surface-altering agent to adhere to the core and/or the density of the surface-altering agent on the core/particle surface. It should be noted that it can be determined by the combination. As such, in some embodiments certain surface modifiers can improve the mobility of one type of particle or core, but not of another type of particle or core.

Although many of the embodiments described herein relate to a single coating, in other embodiments, the particles comprise more than one coating (e.g., at least 2, 3, 4, 5, or Excess coating), and each coating need not be formed of or contain a muco-penetrating material. In some cases, the intermediate coating (ie, the coating between the core surface and the outer coating) may comprise a polymer that facilitates adhesion of the outer coating to the core surface. In many embodiments, the outer coating of the particles comprises a polymer comprising a material that facilitates transport of the particles through the mucus.

As such, the coating (eg, inner coating, intermediate coating, and/or outer coating) may include any suitable polymer. In some cases, the polymer may be biocompatible and/or biodegradable. In some cases, the polymeric material may comprise more than one type of polymer (eg, at least 2, 3, 4, 5, or more polymers). In some cases, the polymer may be a random copolymer or a block copolymer (eg, a diblock copolymer, a triblock copolymer) as described herein.

Non-limiting examples of suitable polymers include polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, polystyrenes, polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyleneimines, Polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylates. Non-limiting examples of specific polymers are poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA) , Poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) ( PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO -Co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacrylate, polyurethane, poly-L-lysine ( PLL), hydroxypropyl methacrylate (HPMA), poly(ethylene glycol), poly-L-glutamic acid, poly(hydroxy acid), polyanhydride, polyorthoester, poly(ester amide), polyamide, polyester Ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly (Ethylene terephthalate), polyvinyl alcohol (PVA), polyvinyl ether, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxane, Polystyrene (PS), polyurethane, derivatized cellulose, such as alkyl cellulose, hydroxyalkyl cellulose, cellulose ether, cellulose ester, nitrocellulose, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acid, such as poly(methyl(meth)) Acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly( Isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly( Isobutyl acrylate), poly(octadecyl acrylate) (collectively referred to herein as “polyacrylic acid”), and copolymers and mixtures thereof, polydioxane and copolymers thereof, polyhydroxyalkanoate, poly Propylene fumarate), polyoxymethylene, poloxamer, poly(ortho)ester, poly(butyric acid), poly(valic acid), poly(lactide-co-caprolactone), and trimethylene carbonate, polyvinylpy Contains lolidone.

The molecular weight of the polymer can be different. In some embodiments, the molecular weight is at least about 0.5 kDa, at least about 1 kDa, at least about 2 kDa, at least about 3 kDa, at least about 4 kDa, at least about 5 kDa, at least about 6 kDa, about 8 kDa, at least about 10 kDa , About 12 kDa or more, about 15 kDa or more, about 20 kDa or more, about 30 kDa or more, about 40 kDa or more, or about 50 kDa or more. In some embodiments, the molecular weight is about 50 kDa or less, about 40 kDa or less, about 30 kDa or less, about 20 kDa or less, about 12 kDa or less, about 10 kDa or less, about 8 kDa or less, about 6 kDa or less, about 5 kDa or less. It may be less than or equal to about 4 kDa. Combinations of the above-mentioned ranges are possible (eg molecular weights of about 2 kDa or more and about 15 kDa or less). Other ranges are also possible. Molecular weight can be determined using any known technique, such as light-scattering and gel permeation chromatography. Other methods are known in the art.

In certain embodiments, the polymer is biocompatible, ie the polymer typically does not induce an adverse response upon insertion or injection into a living body; For example, it does not involve significant inflammation and/or acute rejection of the polymer by the immune system, eg, through a T-cell-mediated response. Of course, it will be appreciated that "biocompatibility" is a relative term, and that some degree of immune response is expected even for polymers that are highly compatible with living tissue. However, as used herein, “biocompatibility” refers to an acute rejection of a substance by at least a portion of the immune system, ie, a non-biocompatible substance implanted in the subject triggers an immune response in the subject, which The substance by is severe enough so that the rejection reaction cannot be adequately controlled, often to the extent that the substance must be removed from the subject. One simple test to measure biocompatibility is exposing the polymer to cells in vitro; Biocompatible polymers are polymers that do not typically cause very little cell death at moderate concentrations, for example at a concentration of about 50 micrograms/10 ⁶ cells. For example, biocompatible polymers can cause less than about 20% cell death upon exposure to cells such as fibroblasts or epithelial cells, even if they are not phagocytized or otherwise uptaken by such cells. In some embodiments, an agent is “biocompatible” if its addition to cells in vitro results in no more than 20% cell death and its administration in vivo does not induce undesired inflammation or other such side effects.

In certain embodiments, the biocompatible polymer may be biodegradable, i.e., the polymer may be chemically and/or biologically (e.g., cellular machinery) within a physiological environment, such as in the body or upon introduction into a cell. ) Or by hydrolysis). For example, the polymer may be one that hydrolyzes spontaneously upon exposure to water (e.g., in a subject) and/or the polymer will decompose upon exposure to heat (e.g., at a temperature of about 37° C.). I can. Decomposition of the polymer can occur at various rates, depending on the polymer or copolymer used. For example, the half-life of the polymer (the time at which 50% of the polymer decomposes into monomeric and/or other non-polymeric moieties) can be on the order of days, weeks, months, or years, depending on the polymer. Polymers can, in some cases, be biologically degraded, for example by exposure to lysozyme (eg, having a relatively low pH), eg by enzymatic activity or cellular machinery. In some cases, the polymer is a monomeric and/or other non-polymeric moiety that can be reused or processed by the cells without a significant toxic effect on the cells (i.e., less than about 20% of the cells are killed when the component is added to the cells in vitro). Can be broken down into tea. For example, polylactide may be hydrolyzed to form lactic acid, polyglycolide may be hydrolyzed to form glycolic acid, and the like.

Examples of biodegradable polymers are poly(ethylene glycol)-poly(propylene oxide)-poly(ethylene glycol) triblock copolymer, poly(lactide) (or poly(lactic acid)), poly(glycolide) (or poly (Glycolic acid)), poly(orthoester), poly(caprolactone), polylysine, poly(ethylene imine), poly(acrylic acid), poly(urethane), poly(anhydride), polyester), poly(trimethylene) Carbonate), poly(ethyleneimine), poly(acrylic acid), poly(urethane), poly(beta amino ester), etc., and copolymers or derivatives of these and/or other polymers, such as poly(lactide- Co-glycolide) (PLGA), but is not limited thereto.

In certain embodiments, the polymer can be biodegradable within a period acceptable for the desired application. In certain embodiments, such as in in vivo therapy, such degradation is usually about 5 years, 1 year, 6 months, 3 months, 1 month, when exposed to a physiological solution having a pH of 6 to 8 at a temperature of 25 to 37°C. It occurs in a period of 15 days, 5 days, less than 3 days, or even 1 day or less (eg 1-4 hours, 4-8 hours, 4-24 hours, 1-24 hours). In other embodiments, the polymer degrades over a period of from about 1 hour to several weeks, depending on the desired application.

Although the coatings and particles described herein may comprise a polymer, in some embodiments, the particles described herein comprise a hydrophobic material that is not a polymer (eg, non-polymeric) and not a pharmaceutical. For example, all or part of the particles may be coated with a passivating layer in some embodiments. Non-limiting examples of non-polymeric materials may include certain metals, waxes, and organic materials (eg, organic silanes, perfluorinated or fluorinated organic materials).

Reduced Particles with mucosal adhesion

As described herein, in some embodiments, the method includes identifying a material, such as a particle, for which it is desirable to reduce its mucoadhesion. Substances requiring increased diffusivity through mucus may, for example, be hydrophobic, have many hydrogen bond donors or acceptors, and/or may be highly charged. In some cases, the material may comprise a crystalline or amorphous solid material. Materials that can serve as cores can be coated with suitable polymers described herein, thereby forming particles with a large number of surface modifiers on the surface, resulting in reduced mucoadhesion. Particles described herein as having reduced mucoadhesion are alternatively characterized as having increased transport through mucus, mobile in mucus, or mucus-penetrating (i.e. mucus-penetrating particles), wherein the particles are (Negative) means that it is transported through the mucus faster than the control particles. The (negative) control particles may be particles known to be mucoadhesive, for example unmodified particles, or cores not coated with a coating described herein, such as 200 nm carboxylated polystyrene particles.

In certain embodiments, the methods herein comprise preparing a pharmaceutical composition or formulation of a modified substance, e.g., with a formulation adapted for delivery (e.g., topical delivery) to a mucus or mucosal surface of a subject. do. Pharmaceutical compositions having a surface-modifying moiety can be delivered to the mucosal surface of a subject, pass through the mucosal barrier in the subject, and/or have prolonged retention, for example due to reduced mucosal adhesion. , The uniform distribution of particles on the mucous membrane surface may be increased. As those skilled in the art will appreciate, mucus is a viscoelastic and sticky substance that captures most exogenous particles. Entrapped particles cannot reach the underlying epithelium and/or are rapidly removed by a mucus clearance mechanism. In order for the particles to reach the underlying epithelium and/or for the particles to prolong their retention in mucosal tissue, the particles must rapidly penetrate mucus secretion and/or avoid mucus clearance mechanisms. If the particles are not substantially attached to the mucus, the particles may diffuse into the interstitial fluid between mucin fibers and reach the underlying epithelium and/or may not be removed by the mucus clearance mechanism. Therefore, modifying a mucoadhesive substance (e.g., a hydrophobic agent) with a substance that reduces mucoadhesion of the particles can enable efficient delivery of the particles to the epithelium of the underlying layer and/or long-term residence on the mucosal surface. have.

Moreover, in some embodiments, particles described herein with reduced mucoadhesion promote better distribution of particles on the tissue surface and/or have a long-term presence on the tissue surface compared to more cohesive particles. For example, in some cases the luminal space, such as the gastrointestinal tract, is surrounded by a mucus-coated surface. Mucoadhesive particles delivered to these spaces are typically removed from the luminal space and from the mucus-coated surface by a natural clearance mechanism in the body. Particles described herein with reduced mucoadhesion may remain in the lumen space for a relatively longer period of time compared to mucoadhesive particles. This long-term presence may prevent or reduce the clearance of the particles and/or allow for better distribution of the particles on the tissue surface. Long-term presence can also affect particle transport through the lumen space, for example particles can be distributed into the mucous layer and reach the underlying epithelium.

In certain embodiments, a material (e.g., a core) coated with a polymer described herein crosses the mucous or mucosal barrier in a subject and/or exhibits prolonged retention and/or increases the uniform distribution of particles on the mucosal surface. For example, such substances are cleared from the subject's body more slowly (e.g., at least 2 times, 5 times, 10 times, or even at least 20 times more slowly) compared to the (negative) control particle. The (negative) control particles may be particles known to be mucoadhesive, for example unmodified particles, or cores not coated with a coating described herein, such as 200 nm carboxylated polystyrene particles.

In certain embodiments, the particles described herein have a specific relative velocity, <V _average > _relative , defined as:

<Equation 1>

In the above equation, <V _mean > is the ensemble average trajectory-mean velocity, V _mean is the individual particle _averaged over its orbit, the sample is the particle of interest, and the negative control is 200 nm carboxylate. Misfired polystyrene particles, positive control is 200 nm polystyrene particles densely PEGylated with 2 kDa-5 kDa PEG.

Relative velocity can be measured by multiple particle tracking techniques. For example, using a fluorescence microscope equipped with a CCD camera, 66.7 ms (15 frames/s) at 100x magnification from several areas in each sample for each type of particle, i.e., sample, negative control, and positive control. It is possible to capture a 15 s movie at a temporal resolution of. Samples, negative and positive controls can be fluorescent particles to observe traces. Alternatively, non-fluorescent particles can be coated with fluorescent molecules, fluorescently tagged surface agents or fluorescently tagged polymers. Measure individual trajectories of multiple particles over a time-scale of 3.335 s (50 frames) or more using advanced image processing software (e.g. Image Pro or MetaMorph) can do.

In some embodiments, the particles described herein are at least about 0.3, at least about 0.4, at least about 0.5, at least about 0.6, at least about 0.7, at least about 0.8, at least about 0.9, at least about 1.0, at least about 1.1, about 1.2 in mucus. At least about 1.3, at least about 1.4, at least about 1.5, at least about 1.6, at least about 1.7, at least about 1.8, at least about 1.9, or at least about 2.0. In some embodiments, the particles described herein are about 10.0 or less, about 8.0 or less, about 6.0 or less, about 4.0 or less, about 3.0 or less, about 2.0 or less, about 1.9 or less, about 1.8 or less, about 1.7 or less, about 1.6 in mucus. Or less, about 1.5 or less, about 1.4 or less, about 1.3 or less, about 1.2 or less, about 1.1 or less, about 1.0 or less, about 0.9 or less, about 0.8 or less, or about 1.7 or less. Combinations of the aforementioned ranges are possible (e.g., relative speeds of about 0.5 to about 6.0). Other ranges are also possible. The mucus can be, for example, mucus of the human cervix.

In certain embodiments, the particles described herein are mucous or mucosal barriers with a greater speed or diffusivity than the control particles or corresponding particles (e.g., corresponding particles that are unmodified and/or not coated with a coating described herein). It can spread through. In some cases, the particles described herein are at least about 10 times, 20 times, 30 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 2000 times, 5000 times, 10000 times than the control particles or corresponding particles. It can cross the mucous or mucosal barrier at a double or even higher diffusion rate. In some cases, the particles described herein are about 10000 times or less, about 5000 times or less, about 2000 times or less, about 1000 times or less, about 500 times or less higher than the control particles or corresponding particles. , Mucus or mucous membrane barrier at a rate of diffusion as high as about 200 times or less, as high as about 100 times, as high as about 50 times, as high as about 30 times, as high as about 20 times, or as high as about 10 times Can pass. Combinations of the above-mentioned ranges are also possible (e.g., about 10 times or more and about 1000 times higher than the control particles or corresponding particles). Other ranges are also possible.

For the purposes of comparison described herein, the corresponding particles may be approximately the same size, shape, and/or density as the test particles, but lack a coating that renders the test particles mobile in the mucus. In some cases, measurements are based on a time scale of about 1 second, or about 0.5 seconds, or about 2 seconds, or about 5 seconds, or about 10 seconds. One skilled in the art will know how to measure the geometric mean square displacement and diffusion rate.

In addition, the particles described herein are at least about 10 times, 20 times, 30 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 2000 times, 5000 times, 10000 times, or It can cross the mucous or mucosal barrier with a higher geometric mean squared displacement. In some cases, the particles described herein are about 10000 times or less, about 5000 times or less, about 2000 times or less, about 1000 times or less, about 500 times or less higher than the control particles or corresponding particles. , A slime with a geometric mean square displacement as high as about 200 times or less, as high as about 100 times, as high as about 50 times, as high as about 30 times, as high as about 20 times or less, or as high as about 10 times or less It can cross the mucous barrier. Combinations of the above-mentioned ranges are also possible (e.g., about 10 times or more and about 1000 times higher than the control particles or corresponding particles). Other ranges are also possible.

In some embodiments, the particles described herein diffuse through the mucosal barrier at a rate that approaches the degree of or rate at which the particles can diffuse through water. In some cases, the particles described herein are about 1/100 or less, about 1/200 or less, about 1/300 or less, about 1/400 or less, about 1/500 or less of the diffusivity at which the particles diffuse through water under the same conditions. , A speed of about 1/600 or less, about 1/700 or less, about 1/800 or less, about 1/900 or less, about 1/1000 or less, about 1/2000 or less, about 1/5000 or less, about 1/10,000 or less, or Diffusion can also cross the mucosal barrier. In some cases, the particles described herein are at least about 1/10,000, at least about 1/5000, at least about 1/2000, at least about 1/1000, at least about 1/900 of the diffusivity at which the particles diffuse through water under the same conditions. , A speed of about 1/800 or more, about 1/700 or more, about 1/600 or more, about 1/500 or more, about 1/400 or more, about 1/300 or more, about 1/200 or more, about 1/100 or more, or Diffusion can also cross the mucosal barrier. Combinations of the aforementioned ranges are also possible (for example, about 1/5000 or more and less than 1/500 of the diffusivity at which the particles diffuse through water under the same conditions). Other ranges are also possible. Measurements can be based on a time scale of about 1 second, or about 0.5 seconds, or about 2 seconds, or about 5 seconds, or about 10 seconds.

In certain embodiments, the particles described herein are capable of diffusing through the mucus of the human cervix with a diffusivity that is less than about 1/500 of the diffusivity at which the particles diffuse through water. In some cases, measurements are based on a time scale of about 1 second, or about 0.5 seconds, or about 2 seconds, or about 5 seconds, or about 10 seconds.

In certain embodiments, the invention provides particles that move through mucus, such as mucus of the human cervix, with a certain degree of absolute diffusivity. For example, the particles described herein may be at least about 1 x 10 ^-4 µm/s, 2 x 10 ^-4 µm/s, 5 x 10 ^-4 µm/s, 1 x 10 ^-3 µm/s, 2 x 10 ^-3 µm/s, 5 x 10 ^-3 µm/s, 1 x 10 ^-2 µm/s, 2 x 10 ^-2 µm/s, 4 x 10 ^-2 µm/s, 5 x 10 ^-2 µm/s , 6 x 10 ^-2 µm/s, 8 x 10 ^-2 µm/s, 1 x 10 ^-1 µm/s, 2 x 10 ^-1 µm/s, 5 x 10 ^-1 µm/s, 1 µm/s , Or a diffusivity of 2 μm/s. In some cases, the particles are about 2 μm/s or less, about 1 μm/s or less, about 5 x 10 ^-1 μm/s or less, about 2 x 10 ^-1 μm/s or less, about 1 x 10 ^-1 μm/s or less. s or less, about 8 x 10 ^-2 µm/s or less, about 6 x 10 ^-2 µm/s or less, about 5 x 10 ^-2 µm/s or less, about 4 x 10 ^-2 µm/s or less, about 2 x 10 ^-2 µm/s or less, about 1 x 10 ^-2 µm/s or less, about 5 x 10 ^-3 µm/s or less, about 2 x 10 ^-3 µm/s or less, about 1 x 10 ^-3 µm/s Hereinafter, a diffusion degree of about 5 x 10 ^-4 µm/s or less, about 2 x 10 ^-4 µm/s or less, or about 1 x 10 ^-4 µm/s or less may be moved. Combinations of the above mentioned ranges are also possible (eg, greater than or equal to about 2×10 ⁻⁴ μm/s and less than or equal to about 1×10 ⁻¹ μm/s). Other ranges are also possible. In some cases, measurements are based on a time scale of about 1 second, or about 0.5 seconds, or about 2 seconds, or about 5 seconds, or about 10 seconds.

While many of the mobility (eg, relative velocity, diffusivity) described herein can be measured in mucus of the human cervix, it should be noted that they can likewise be measured in other types of mucus.

In certain embodiments, the particles described herein comprise surface modifying moieties at a given density. The surface modifying moiety can be a portion of the surface modifying agent that is exposed to, for example, a solvent containing the particles. As an example, the hydrolyzed units/block of PVA may be the surface modifying moiety of the surface modifying agent PVA. In some cases, the surface modifying moiety and/or surface modifying agent is about 0.001 or more units or molecules per nm2, about 0.002 or more, about 0.005 or more, about 0.01 or more, about 0.02 or more, about 0.05 or more, about 0.1 At least about 0.2, at least about 0.5, at least about 1, at least about 2, at least about 5, at least about 10, at least about 20, at least about 50, at least about 100 units or molecules, or more units per nm 2 or It exists in the density of molecules. In some cases, the surface modifying moiety and/or surface modifying agent is about 100 or less units or molecules per nm2, about 50 or less, about 20 or less, about 10 or less, about 5 or less, about 2 or less, about 1 or less, about 0.5 or less, about 0.2 or less, about 0.1 or less, about 0.05 or less, about 0.02 or less, or about 0.01 or less units or molecules. Combinations of the above-mentioned ranges are also possible (eg, density of about 0.01 or more and about 1 unit or molecule per nm 2 ). Other ranges are also possible. In some embodiments, the density value described above may be an average density because the surface modifier is in equilibrium with other components in the solution.

One of skill in the art will know how to estimate the average density of the surface modifying moiety (see, for example, SJ Budijono et al., Colloids and Surfaces A: Physicochem. Eng. Aspects 360 (2010) 105-110) and [Joshi]. , et al ., Anal . Chim . Acta 104 (1979) 153-160, each of which is incorporated herein by reference). For example, as described herein, the average density of surface modifying moieties can be determined using HPLC quantification and DLS analysis. A suspension of particles of interest for surface density determination is first sized using DLS: a small volume is diluted to an appropriate concentration (e.g. -100 μg/mL) and the z-average diameter is taken as a representative measure of the particle size. do. The remaining suspension is then divided into two aliquots. Using HPLC, the first aliquot is assayed for the total concentration of the core material and the total concentration of the surface modifying moiety. Again HPLC is used to assay the second aliquot for the concentration of free or unbound surface modifying moieties. In order to obtain only free or unbound surface modifying moieties from the second aliquot, the particles, and thus any bound surface modifying moieties, are removed by ultracentrifugation. By subtracting the concentration of unbound surface modifying moieties from the total concentration of surface modifying moieties, the concentration of bound surface modifying moieties can be determined. Further, since the total concentration of the core material was determined from the first aliquot, the mass ratio of the core material and the surface modifying moiety can be determined. The molecular weight of the surface modifying moiety can be used to calculate the number of surface modifying moieties relative to the mass of the core material. To convert this number into a surface density measurement, it is necessary to calculate the surface area per mass of the core material. The volume of the particles is calculated as an approximation as the volume of a sphere having a diameter obtained from DLS, making it possible to calculate the surface area per mass of the core material. In this way the number of surface modifying moieties per surface area can be determined. Unless otherwise specified, density values herein were determined using this method.

In certain embodiments, the particles described herein may include surface modifying moieties and/or surface modifying agents that affect the zeta potential of the particles. The zeta potential of the coated particles is, for example, about -100 mV or more, about -75 mV or more, about -50 mV or more, about -40 mV or more, about -30 mV or more, about -20 mV or more, about -10. mV or more, about -5 mV or more, about 5 mV or more, about 10 mV or more, about 20 mV or more, about 30 mV or more, about 40 mV or more, about 50 mV or more, about 75 mV or more, or about 100 mV or more . Combinations of the above-mentioned ranges are possible (eg, a zeta potential of about -50 mV or more and about 50 mV or less). Other ranges are also possible.

The coated particles described herein can have any suitable shape and/or size. In some embodiments, the coated particles have a shape substantially similar to that of the core. In some cases, the coated particles described herein may be nanoparticles, i.e., the particles have a characteristic dimension of less than about 1 micron, wherein the characteristic dimension of the particle is the diameter of a full sphere having the same volume as the particle. In other embodiments, larger sizes are possible (eg, about 1-10 microns). The plurality of particles can also be characterized by an average size (eg, an average maximum cross-sectional dimension, or an average minimum cross-sectional dimension for the plurality of particles), in some embodiments. Multiple particles are, for example, about 10 μm or less, about 5 μm or less, about 1 μm or less, about 800 nm or less, about 700 nm or less, about 500 nm or less, 400 nm or less, 300 nm or less, about 200 nm or less , About 100 nm or less, about 75 nm or less, about 50 nm or less, about 40 nm or less, about 35 nm or less, about 30 nm or less, about 25 nm or less, about 20 nm or less, about 15 nm or less, or about 5 nm or less Can have an average size of In some cases, the plurality of particles may be, for example, about 5 nm or more, about 20 nm or more, about 50 nm or more, about 100 nm or more, about 200 nm or more, about 300 nm or more, about 400 nm or more, about 500 nm. It may have an average size of about 1 μm or more and about 5 μm or more. Combinations of the above mentioned ranges are also possible (e.g., an average size of about 50 nm or more and about 500 nm or less). Other ranges are also possible. In some embodiments, the size of the core formed by the methods described herein has a Gaussian distribution.

drugs

In some embodiments, the coated particles comprise one or more agents. The medicament may be present in the core of the particle and/or in the coating of the particle (eg, dispersed throughout the core and/or coating). In some cases, the medicament may be disposed on the surface of the particle (eg, on the outer surface of the coating, on the inner surface of the coating, on the surface of the core). The medicament can often be contained within the particle and/or placed in a portion of the particle using known techniques (eg, coating, adsorption, covalent linkage, encapsulation, or other process). In some cases, the agent may be present in the core of the particle prior to or during coating of the particle. In some cases, the agent is present during formation of the core of the particle, as described herein.

Non-limiting examples of medicaments include imaging agents, diagnostic agents, therapeutic agents, agents with detectable labels, nucleic acids, nucleic acid analogs, small molecules, peptidomimetics, proteins, peptides, lipids, vaccines, viral vectors, viruses, and surfactants. do.

In some embodiments, the agent contained in the particles described herein has a therapeutic, diagnostic, or imaging effect in the targeted mucosal tissue. Non-limiting examples of mucosal tissues include oral cavity (including, for example, buccal and esophageal membranes and tonsil surfaces), eyes, gastrointestinal tract (including, for example, stomach, small intestine, large intestine, colon, rectum), nasal cavity, respiratory tract (for example, , Nasal cavity, pharynx, trachea and bronchial membranes), and genital (eg, vaginal, cervical and urethral membranes) tissues.

Any suitable number of medicaments can be present in the particles described herein. For example, at least 1, at least 2, at least 3, at least 4, at least 5, or more, but generally less than 10 agents may be present in the particles described herein.

The number of drugs that are mucoadhesive is known in the art and can be used as a medicament in the particles described herein (e.g., Khanvilkar K, Donovan MD, Flanagan DR, Drug transfer through mucus, Advanced Drug Delivery Reviews 48 (2001) 173-193]; [Bhat PG, Flanagan DR, Donovan MD. Drug diffusion through cystic fibrotic mucus: steady-state permeation, rheologic properties, and glycoprotein morphology, J Pharm Sci, 1996 Jun;85(6):624 -30]). Further non-limiting examples of medicaments include imaging and diagnostic agents (such as radiopaque agents, labeled antibodies, labeled nucleic acid probes, dyes such as colored or fluorescent dyes, etc.) and adjuvants (radiation sensitizers, transfection enhancers, Including chemotactic agents and chemoattractants, peptides that modulate cell adhesion and/or cell mobility, cell penetrants, vaccine potentiators, multidrug resistance and/or inhibitors of efflux pumps, etc.) do.

Further non-limiting examples of pharmaceuticals include aoxyprine, auranopine, azapropazone, verorilate, diflunisal, etodolac, fenbufen, fenoprofen calsim, flurbiprofen, furosemide, ibuprofen. , Indomethacin, ketoprofen, loteprednol etabonate, meclofenamic acid, mefenamic acid, nabumetone, naproxen, oxyfenbutazone, phenylbutazone, piroxen, sulindac, albendazole, bee Phenium Hydroxynaphthoate, Cambendazole, Dichlorophene, Ivermectin, Mebendazole, Oxaniquine, Oxpendazole, Oxantel Embonate, Praziquantel, Pyrantel Embonate, Thiabendazole, Amiodarone HCl , Disopyramide, flecainide acetate, quinidine sulfate. Antibacterial agents: Benetamine penicillin, sinoxacin, ciprofloxacin HCl, clarithromycin, clofazimine, cloxacillin, demeclocycline, doxycycline, erythromycin, ethionamide, imipenem, nalidic acid, nitrofuran Toin, rifampicin, spiramycin, sulfabenzamide, sulfadoxine, sulfamerazine, sulfacetamide, sulfadiazine, sulfafurazole, sulfametoxazole, sulfapyridine, tetracycline, trimethoprim, dicumarol, diffi Lidamol, Nicumalon, Penindione, Amoxapine, Maprotiline HCl, Myanserine HCL, Nortriptyline HCl, Trazodone HCL, Trimipramine Maleate, Acetohexamide, Chlorpropamide, Gleebenclamide , Glyclazide, Glipizide, Tolazamide, Tolbutamide, Beclamide, Carbamazepine, Clonazepam, Etotoin, Metoin, Metsuccivid, Methylphenobarbitone, Oxcarbazepine , Paramethadione, phenasemide, phenobarbitone, phenytoin, pensuccimide, primidone, sultiam, valproic acid, amphotericin, butoconazole nitrate, clotrimazole, econasol nitrate, fluconazole, flu Cytosine, griseofulvin, itraconazole, ketoconazole, miconazole, natamycin, nystatin, sulconazole nitrate, terbinafine HCl, terconazole, thioconazole, undecenoic acid, allopurinol, propeneside, sulfin -Pyrazone, amlodipine, benidipine, darodipine, dilitagem HCl, diazoxide, felodipine, guanabenz acetate, isradipine, minoxidil, nicardipine HCl, nifedipine, nimodipine, phenoxybenzamine HCl , Prazosin HCL, reserpine, terazosin HCL, amodiaquine, chloroquine, chlorproguanil HCl, halopanthrin HCl, mefloquine HCl, roguanyl HCl, pyrimethamine, quinine sulfate, dihydroergotamine mesyl Rate, Ergotamine Tartrate, Methicergide Maleate, Pizotifen Maleate, Sumatriptan Succinate, Atropine, Benzhexol HCl, Biperidene, Etopropazine HCl, Hioxiamine, Mefenzolate Bromide, Oxyphencyclamine HCl, tropicamide, aminoglutetimide, amsacrine, azathioprine, bu Sulfan, Chlorambucil, Cyclosporine, Dacarbazine, Estramustine, Etoposide, Lomustine, Melphalan, Mercaptopurine, Methotrexate, Mitomycin, Mitophan, Mitoxantrone, Procarbazine HCl, Tamoxifen Citrate , Testolactone, benznidazole, clioquinol, decoquinate, diiodohydroxyquinoline, diroxanide furoate, dinitolmid, purzolidone, metronidazole, nimorazole, nitrofurazone, ornidazole, tinida Sol, carbimazole, propylthiouracil, alprazolam, amylobarbitone, barbitone, bentazepam, bromazepam, bromperidol, brotizolam, butobarbitone, carbromal, chlordiazepoxide, Chlormethiazole, chlorpromazine, clobazam, clothiazepam, clozapine, diazepam, droperidol, etinamate, pullunanisone, flunitrazepam, fluopramazine, flupentixol decanoate, flufenazine de Canoate, flurazepam, haloperidol, lorazepam, lormethazepam, medazepam, meprobamate, metaqualon, midazolam, nitrazepam, oxazepam, pentobarbitone, perphenazine pimozide, prochlorperazine, Sulpiride, Temazepam, Thioridazine, Triazolam, Zopiclone, Acebutolol, Alprenolol, Atenolol, Labetalol, Metoprolol, Nadolol, Oxprenolol, Pindolol, Propanolol, Amlinone, Digitoxin , Digoxin, Enoxymon, Ranatoside C, Medigoxin, Beclomethasone, Betamethasone, Budesonide, Cortisone Acetate, Desoxymethasone, Dexamethasone, Fludrocortisone Acetate, Flunisolide, Flucortolone, Fluticasone Pro Cypionate, hydrocortisone, methylprednisolone, prednisolone, prednisone, traamcinolone, acetazolamide, amiloride, bendrofluazide, bumetanide, chlorothiazide, chlorthalidone, ethacrynic acid, prusce Mid, metolazone, spironolactone, triamterene, bromocriptine mesylate, lisuride maleate, bisacodyl, cimetidine, cisapride, diphenoxylate HCl, domperidone, famotidine, loperamide, mesala Gin, Nizatidine, Omeprazole, Ondansetron HCL, Ranitidine HCl, Sulfasalazine, Acri Bastine, Astemizol, Cinnarizine, Cyclizine, Ciproheptadiene HCl, Dimenhydrinate, Pullunarizine HCl, Loratadine, Meclozin HCl, Oxatomide, Terfenadine, Bennafibrate, Clofibrate , Fenofibrate, gemfibrozil, probucol, amyl nitrate, glyceryl trinitrate, isosorbide dinitrate, isosorbide mononitrate, pentaerythritol tetranitrate, beta carotene, vitamin A, vitamin B2, Vitamin D, vitamin E, vitamin K, codeine, dextropropoxyfen, diamorphine, dihydrocodeine, methazinol, methadone, morphine, nalbuphine, pentazosin, clomiphene citrate, danazole, ethinyl estradiol, medroxy Progesterone acetate, mestranol, methyltestosterone, norethysterone, norgestrel, estradiol, conjugated estrogen, progesterone, stanozolol, stivestrol, testosterone, tibolone, amphetamine, dexamphetamine, dexfenfluramine, fenfluramine, And margin stone.

Uses and pharmaceutical compositions

The particles described herein can be used in any suitable application. In some cases, the particle is a part of the pharmaceutical composition (e.g., as described herein), e.g., through or to a mucous or mucous membrane surface, to which an agent (e.g., drug, therapeutic agent, diagnostic agent, imaging agent) It is used to convey. Pharmaceutical compositions may comprise one or more particles described herein and one or more pharmaceutically acceptable excipients or carriers. The composition can be used to treat, prevent, and/or diagnose a condition in a subject, wherein the method comprises administering a pharmaceutical composition to the subject. Subjects or patients treated by the articles and methods described herein can refer to humans or non-human animals such as primates, mammals, and vertebrates.

Methods related to treating a subject include preventing the disease, disorder, or condition from occurring in a subject that may be susceptible to a disease, disorder, and/or condition but has not yet been diagnosed with the disease; Inhibiting a disease, disorder or condition, eg, delaying its progression; And alleviating the disease, disorder, or condition, eg, causing regression of the disease, disorder and/or condition. Treating a disease or condition involves ameliorating one or more symptoms of a particular disease or condition, even if the underlying pathophysiology is not affected (e.g., such analgesic drugs do not treat the cause of pain. Such treatment of pain in the subject by administering).

In some embodiments, the pharmaceutical compositions described herein can be delivered to the mucosal surface of a subject and pass through the mucosal barrier (e.g., mucus) in the subject and/or, due to, for example, reduced mucosal adhesion, and/or May exhibit prolonged retention and/or increased uniform distribution of the particles in. Non-limiting examples of mucosal tissues include oral cavity (including, for example, buccal and esophageal membranes and tonsil surfaces), eyes, gastrointestinal tract (including, for example, stomach, small intestine, large intestine, colon, rectum), nasal cavity, respiratory tract (for example, , Nasal cavity, pharynx, trachea and bronchial membranes), genitals (eg, vaginal, cervical and urethral membranes).

Pharmaceutical compositions described herein and for use in accordance with the articles and methods described herein may comprise a pharmaceutically acceptable excipient or carrier. Pharmaceutically acceptable excipients or pharmaceutically acceptable carriers may comprise any suitable type of non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid. Some examples of substances that can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; Starch, such as corn starch and potato starch; Cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; Powdered tragacanth; malt; gelatin; Talc; Excipients such as cocoa butter and suppository wax; Oil, peanut oil, cottonseed oil; Safflower oil; Sesame oil; olive oil; Corn oil and soybean oil; Glycols such as propylene glycol; Esters such as ethyl oleate and ethyl laurate; Agar; Detergents such as Tween 80; Buffering agents such as magnesium hydroxide and aluminum hydroxide; Alginic acid; Pyrogen-free water; Isotonic saline; Ringer's solution; Ethyl alcohol; And phosphate buffers, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening agents, flavoring agents and fragrances, preservatives and antioxidants, as well as at the discretion of the formulator. Can be present in the composition. As will be appreciated by those of skill in the art, excipients may be selected based on the route of administration, the medicament delivered, the time course of delivery of the agent, and the like, as described below.

Pharmaceutical compositions containing the particles described herein can be administered to a subject via any route known in the art. These are oral, sublingual, nasal, intradermal, subcutaneous, intramuscular, rectal, vaginal, intravenous, intraarterial, intravenous, intraperitoneal, intravitreal, periorbital, topical (as a powder, cream, ointment, or drop), Includes but is not limited to buccal and inhalation administration. In some embodiments, the compositions described herein may be administered parenterally as an injection (intravenous, intramuscular, or subcutaneous), drop infusion formulation, or suppository. As will be appreciated by those skilled in the art, the route of administration and effective dosage that achieves the desired biological effect will be determined by the agent to be administered, the target organ, the agent to be administered, the time course of administration, the disease to be treated, the intended use, etc. I can.

As an example, the particles can be included in a pharmaceutical composition formulated as a nasal spray such that the pharmaceutical composition is delivered over the mucous layer of the nose. As another example, the particles can be included in a pharmaceutical composition formulated as an inhaler such that the pharmaceutical composition is delivered across the mucous layer of the lungs. As another example, when the composition is administered orally, it may be formulated as a tablet, capsule, granule, powder, or syrup. Similarly, the particles can be included in pharmaceutical compositions that are delivered through the eye, gastrointestinal tract, nasal cavity, respiratory tract, rectum, urethra and/vaginal tissue.

For application by the ophthalmic mucosal route, the subject composition may be formulated as an eye drop or eye ointment. These formulations can be prepared in a conventional manner and, if desired, the composition of interest to be mixed with any conventional additives such as buffers or pH-adjusters, tonicity modifiers, viscosity modifiers, suspension stabilizers, preservatives, and other pharmaceutical excipients. I can. Moreover, in certain embodiments, the subject compositions described herein may be lyophilized or subjected to another suitable drying technique, such as spray drying.

In some embodiments, the particles described herein, which can be administered in an inhaled or aerosol formulation, comprise one or more agents useful in inhalation therapy, such as adjuvants, diagnostic agents, imaging agents, or therapeutic agents. The particle size of the particulate medicament should be such that substantially all of the medicament can be inhaled into the lungs upon administration of the aerosol formulation, e.g., less than about 20 microns, such as about 1 to about 10 microns, for example about It may range from 1 to about 5 microns, but other ranges are also possible. The particle size of the medicament can be reduced in a conventional manner, for example by milling or micronisation. Alternatively, the particulate medicament can be administered to the lungs by spraying of a suspension. The final aerosol formulation contains, for example, a medicament of 0.005-90% w/w, 0.005-50%, 0.005-10%, about 0.005-5% w/w, or 0.01-1.0% w/w relative to the total amount of the formulation. can do. Other ranges are also possible.

While never required, it is preferred that the formulations described herein do not contain any ingredients that may cause decomposition of stratospheric ozone. In particular, in some embodiments, a propellant is selected that does not contain or does not consist essentially of chlorofluorocarbons such as CCl ₃ F, CCl ₂ F ₂ , and CF ₃ CCl ₃ .

Aerosols may contain propellants. The propellant may optionally contain an adjuvant having a higher polarity and/or a higher boiling point than the propellant. Polarity, which can be used to adjuvants (e. G., C _{2 -6)} aliphatic alcohols and polyols such as ethanol, isopropanol, and propylene glycol, preferably ethanol. In general, only a small amount of polar adjuvant (e.g., 0.05-3.0% w/w) may be needed to improve the stability of the dispersion, and the use of an amount greater than 5% w/w tends to dissolve the drug. This can be. Formulations according to embodiments described herein may contain less than 1% w/w, for example about 0.1% w/w, of a polar adjuvant. However, the formulations described herein may be substantially free of polar adjuvants, especially ethanol. Suitable volatile adjuvants include saturated hydrocarbons such as propane, n-butane, isobutane, pentane and isopentane and alkyl ethers such as dimethyl ether. In general, up to 50% w/w of the propellant may comprise up to 30% w/w of a volatile adjuvant, for example a volatile saturated C ₁ -C ₆ hydrocarbon. Optionally, the aerosol formulation according to the invention may further comprise one or more surfactants. Surfactants may be physiologically acceptable when administered by inhalation. Surfactants within this category, such as L-α-phosphatidylcholine (PC), 1,2-dipalmitoylphosphatidylcholine (DPPC), oleic acid, sorbitan trioleate, sorbitan mono-oleate, sorbitan monolaurate, poly Oxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, natural lecithin, oleyl polyoxyethylene ether, stearyl polyoxyethylene ether, lauryl polyoxyethylene ether, block copolymer of oxyethylene and oxypropylene , Synthetic lecithin, diethylene glycol dioleate, tetrahydrofurfuryl oleate, ethyl oleate, isopropyl myristate, glyceryl monooleate, glyceryl monostearate, glyceryl monoricinolate, cetyl alcohol, ste Aryl alcohol, polyethylene glycol 400, cetyl pyridinium chloride, benzalkonium chloride, olive oil, glyceryl monolaurate, corn oil, cottonseed oil, and sunflower seed oil.

The formulations described herein can be prepared by dispersion of the particles in a selected propellant and/or co-propellant in an appropriate container, eg utilizing sonication. The particles can be suspended in a co-propellant and filled into a suitable container. The valve of the container is then sealed in place and the propellant is introduced by pressure filling through the valve in a conventional manner. Thus, the particles can be suspended or dissolved in a liquefied propellant, sealed in a container equipped with a metering valve, and fitted into an actuator. Such measured dose inhalers are well known in the art. The metering valve can meter 10 to 500 μL, preferably 25 to 150 μL. In certain embodiments, dispersion can be achieved using a dry powder inhaler (eg, a spinhaler) for the particles (remaining as dry powder). In other embodiments, nanospheres can be aerosolized into the lungs by being suspended in an aqueous fluid and sprayed into fine droplets.

Ultrasonic nebulizers can be used because they minimize exposure of the agent to shear, which can lead to particle decomposition. Usually, aqueous aerosols are prepared by formulating an aqueous solution or suspension of particles with conventional pharmaceutically acceptable carriers and stabilizers. Carriers and stabilizers will depend on the particular composition requirement, typically a non-ionic surface active agent (Tween, Pluronic ^®, or polyethylene glycol), innocuous proteins such as serum albumin, sorbitan esters, oleic acid, lecithin, amino acids , Such as glycine, buffers, salts, sugars, or sugar alcohols. Aerosols are generally prepared from isotonic solutions.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and exisirs. In addition to the active ingredients (i.e., microparticles, nanoparticles, liposomes, micelles, polynucleotide/lipid complexes), liquid dosage forms can be used with inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, For example ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, Germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, fatty acid esters of polyethylene glycol and sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants such as emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Formulations for injection, for example sterile aqueous or oily suspensions for injection, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be as sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, for example as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, U.S.P. And isotonic sodium chloride solution. In addition, sterile, fixed oils are commonly used as solvents or suspension media. Any bland fixed oil can be used for this purpose including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the manufacture of injections. In certain embodiments, the particles are suspended in a carrier fluid comprising 1% (w/v) sodium carboxymethyl cellulose and 0.1% (v/v) Tween 80.

Formulations for injection may be sterilized, for example, by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use, or by filtration through a bacteria-retaining filter. I can.

Compositions for rectal or vaginal administration can be suppositories that can be prepared by mixing the particles with a suitable bland excipient or carrier, such as cocoa butter, polyethylene glycol, or suppository wax, which is solid at ambient temperature but liquid at body temperature and thus rectal Or it melts in the vaginal cavity and releases particles.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the particles may contain one or more inert, pharmaceutically acceptable excipients or carriers such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starch, lactose, sucrose, glucose, Mannitol and silicic acid, b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, etc., c) wetting agents such as glycerol, d) disintegrants such as agar-agar, Calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl Alcohol and glycerol monostearate and the like, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. . In the case of capsules, tablets and pills, the dosage form may also include a buffering agent.

Solid compositions of a similar type can also be used as fillers in soft and hard-filled gelatin capsules using high molecular weight polyethylene glycols and the like as well as excipients such as lactose or lactose.

Solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They may optionally contain opacifying agents and may also be compositions that release only the active ingredient(s), or preferentially, in a certain part of the intestine, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric materials and waxes.

Solid compositions of a similar type can also be used as fillers in soft and hard-filled gelatin capsules using high molecular weight polyethylene glycols and the like as well as excipients such as lactose or lactose.

Dosage forms for topical or transdermal administration of the pharmaceutical composition of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The particles are mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. Ophthalmic formulations, ear drops, and eye drops are also contemplated as being within the scope of the present invention.

Ointments, pastes, creams, and gels, in addition to the particles described herein, include excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, tragacanths, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and Zinc oxide, or mixtures thereof.

Powders and sprays may contain, in addition to the particles described herein, excipients such as lactose, talc, silicic acid, aluminum oxide, calcium silicate, and polyamide powder, or mixtures of these substances. Sprays may also contain customary propellants, such as chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlled delivery of compounds to the body. Such dosage forms can be prepared by dissolving or dispersing the microparticles or nanoparticles in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound across the skin. The rate can be controlled by providing a rate controlling membrane or by dispersing the particles in a polymer matrix or gel.

The particles described herein, including the medicament, can be administered to a subject in an amount sufficient to deliver a therapeutically effective amount of the incorporated medicament to the subject as part of a diagnostic, prophylactic, or therapeutic treatment. In general, an effective amount of a drug or ingredient refers to the amount required to elicit the desired biological response. The desired concentration of the drug in the particles includes, but is not limited to, the rate of absorption, inactivation, and excretion of the drug, as well as the rate of delivery of the compound from the subject composition, the desired biological endpoint, the agent to be delivered, the target tissue, etc. Will be determined by It should be noted that the dosage value may also vary depending on the severity of the condition being alleviated. In the case of any particular subject, it should be further understood that the particular dosage regimen should be adjusted over time according to the individual needs and the professional judgment of the person administering or supervising the administration of the composition. Typically, administration will be determined using techniques known to those of skill in the art.

The medicament may be in any suitable amount, for example, at least about 0.01 wt%, at least about 0.1 wt%, at least about 1 wt%, at least about 5 wt%, at least about 10 wt%, about 20 wt% of the composition and/or formulation. It may be present in the composition and/or formulation as above. In some cases, the agent may be present in the composition and/or formulation at about 30 wt% or less, about 20 wt% or less, about 10 wt% or less, about 5 wt% or less, about 2 wt% or less, or about 1 wt% or less. have. Combinations of the aforementioned ranges are also possible (eg present in an amount of about 0.1 wt% or more and about 10 wt% or less). Other ranges are also possible. In certain embodiments, the pharmaceutical agent is about 0.1-2 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 2-20 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 0.2 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 0.4 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 1 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 2 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 5 wt% of the composition and/or formulation. In certain embodiments, the pharmaceutical agent is about 10 wt% of the composition and/or formulation.

The concentration and/or amount of any medicament administered to a subject can be readily determined by one of skill in the art. Known methods are also available for assaying local tissue concentration, rate of diffusion from particles, and local blood flow before and after administration of a therapeutic agent.

The compositions and/or formulations described herein may have any suitable osmolarity. In some embodiments, the compositions and/or formulations described herein are at least about 0 mOsm/L, at least about 5 mOsm/L, at least about 25 mOsm/L, at least about 50 mOsm/L, at least about 75 mOsm/L, about It may have an osmolarity of 100 mOsm/L or more, about 150 mOsm/L or more, about 200 mOsm/L or more, about 250 mOsm/L or more, or about 310 mOsm/L or more. In certain embodiments, the compositions and/or formulations described herein are about 310 mOsm/L or less, about 250 mOsm/L or less, about 200 mOsm/L or less, about 150 mOsm/L or less, about 100 mOsm/L or less, about It may have an osmolarity of 75 mOsm/L or less, about 50 mOsm/L or less, about 25 mOsm/L or less, or about 5 mOsm/L or less. Combinations of the above mentioned ranges are also possible (eg osmolarity of about 0 mOsm/L or more and about 50 mOsm/L or less). Other ranges are also possible. The osmolarity of the composition and/or formulation can be varied, for example, by changing the concentration of the salt present in the solvent of the composition and/or formulation.

In one set of embodiments, the composition and/or formulation comprises one or more chelating agents. As used herein, chelating agents refer to chemical compounds that have the ability to react with metal ions to form complexes through one or more bonds. The one or more bonds are typically ionic or coordination bonds. The chelating agent can be an inorganic or organic compound. Metal ions capable of catalyzing certain chemical reactions (eg, oxidation reactions) may lose their catalytic activity when the metal ions bind to a chelating agent to form a complex. Thus, the chelating agent can exhibit preservative properties when it binds to metal ions. Any suitable chelating agent having preservative properties can be used, such as phosphonic acids, aminocarboxylic acids, hydroxycarboxylic acids, polyamines, aminoalcohols, and polymer chelating agents. Specific examples of the chelating agent include ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), diethylenetriaminepentacetic acid (DTPA), N -hydroxyethylethylene diaminetriacetic acid (HEDTA), tetraborate, triethylamine. Diamines, and salts and derivatives thereof, but are not limited thereto. In certain embodiments, the chelating agent is EDTA. In certain embodiments, the chelating agent is a salt of EDTA. In certain embodiments, the chelating agent is disodium EDTA.

The chelating agent may be present in a suitable concentration in the composition and/or formulation comprising the coated particles described herein. In certain embodiments, the concentration of the chelating agent is at least about 0.0003 wt%, at least about 0.001 wt%, at least about 0.003 wt%, at least about 0.01 wt%, at least about 0.03 wt%, at least about 0.05 wt%, about 0.1 wt% At least about 0.3 wt%, at least about 1 wt%, or at least about 3 wt%. In certain embodiments, the concentration of chelating agent is about 3 wt% or less, about 1 wt% or less, about 0.3 wt% or less, about 0.1 wt% or less, about 0.05 wt% or less, about 0.03 wt% or less, about 0.01 wt% Or less, about 0.003 wt% or less, about 0.001 wt% or less, or about 0.0003 wt% or less. Combinations of the above-mentioned ranges are possible (eg, concentrations of about 0.01 wt% or more and about 0.3 wt% or less). Other ranges are also possible. In certain embodiments, the concentration of chelating agent is about 0.001-0.1 wt%. In certain embodiments, the concentration of chelating agent is about 0.005 wt%. In certain embodiments, the concentration of chelating agent is about 0.01 wt%. In certain embodiments, the concentration of chelating agent is about 0.05 wt%. In certain embodiments, the concentration of chelating agent is about 0.1 wt%.

In some embodiments, the chelating agent may be present in the composition and/or formulation in one or more of the aforementioned ranges during the formation and/or dilution processes described herein. In certain embodiments, the chelating agent may be present in the composition and/or formulation in one or more of the aforementioned ranges in the final product.

In some embodiments, antimicrobial agents may be included in compositions and/or formulations comprising the coated particles described herein. Antimicrobial agents, as used herein, generally refer to bioactive agents effective in inhibiting, preventing, or protecting against microorganisms, such as bacteria, microbes, fungi, viruses, spores, yeasts, fungi, etc., associated with infection. Examples of antimicrobial agents include cephaloporin, clindamycin, chloramphenicol, carbapenem, minocycline, lymphapine, penicillin, monobactam, quinolones, tetracyclines, macrolides, sulfa antibiotics, trimethoprim, fu Nanoparticles of cid acid, aminoglycoside, amphotericin B, azole, flucytosine, silofulgin, salibacterial nitrofuran compound, metallic silver or a silver alloy containing about 2.5 wt% copper, silver citrate, silver acetate, benzoic acid Silver, bismuth pyrithione, zinc pyrithione, zinc percarbonate, zinc perborate, bismuth salts, parabens (e.g. methyl-, ethyl-, propyl-, butyl-, and octyl-benzoic acid esters), citric acid, benzal Conium chloride (BAC), limpamycin, and sodium percarbonate.

Antimicrobial agents may be present in suitable concentrations in compositions and/formulations comprising the coated particles described herein. In certain embodiments, the concentration of the antimicrobial agent is at least about 0.0003 wt%, at least about 0.001 wt%, at least about 0.003 wt%, at least about 0.01 wt%, at least about 0.03 wt%, at least about 0.1 wt%, about 0.3 wt% Or more, about 1 wt% or more, or about 3 wt% or more. In certain embodiments, the concentration of the antimicrobial agent is about 3 wt% or less, about 1 wt% or less, about 0.3 wt% or less, about 0.1 wt% or less, about 0.03 wt% or less, about 0.01 wt% or less, about 0.003 wt% Below, it may be about 0.001 wt% or less, or about 0.0003 wt% or less. Combinations of the above-mentioned ranges are possible (eg, concentrations of about 0.001 wt% or more and about 0.1 wt% or less). Other ranges are also possible. In certain embodiments, the concentration of antimicrobial agent is about 0.001-0.05 wt%. In certain embodiments, the concentration of antimicrobial agent is about 0.002 wt%. In certain embodiments, the concentration of antimicrobial agent is about 0.005 wt%. In certain embodiments, the concentration of antimicrobial agent is about 0.01 wt%. In certain embodiments, the concentration of antimicrobial agent is about 0.02 wt%. In certain embodiments, the concentration of antimicrobial agent is about 0.05 wt%.

In some embodiments, the antimicrobial agent may be present in the composition and/or formulation in one or more of the aforementioned ranges during the formation and/or dilution processes described herein. In certain embodiments, the antimicrobial agent may be present in the composition and/or formulation in one or more of the aforementioned ranges in the final product.

In some embodiments, tonicity agents may be included in compositions and/or formulations comprising the coated particles described herein. As used herein, an isotonic agent refers to a compound or substance that can be used to adjust the composition of a formulation to a desired osmolarity range. In certain embodiments, the desired osmolarity range is an isotonic range compatible with blood. In certain embodiments, the desired osmolarity range is shelf life. In certain embodiments, the desired osmolarity range is hypertonic. Examples of isotonic agents include glycerin, lactose, mannitol, dextrose, sodium chloride, sodium sulfate, sorbitol, salt-sodium citrate (SSC), and the like. In certain embodiments, a combination of one or more tonicity agents may be used. In certain embodiments, the tonicity agent is glycerin. In certain embodiments, the tonicity agent is sodium chloride.

The tonicity agent (such as those described herein) may be present in a suitable concentration in the composition and/or formulation comprising the coated particles described herein. In certain embodiments, the concentration of the tonicity agent is at least about 0.003 wt%, at least about 0.01 wt%, at least about 0.03 wt%, at least about 0.1 wt%, at least about 0.3 wt%, at least about 1 wt%, about 3 wt% At least about 10 wt%, at least about 20 wt%, or at least about 30 wt%. In certain embodiments, the concentration of the tonicity agent is about 30 wt% or less, about 10 wt% or less, about 3 wt% or less, about 1 wt% or less, about 0.3 wt% or less, about 0.1 wt% or less, about 0.03 wt% Or less, about 0.01 wt% or less, or about 0.003 wt% or less. Combinations of the above-mentioned ranges are possible (eg, concentrations of about 0.1 wt% or more and about 10 wt% or less). Other ranges are also possible. In certain embodiments, the concentration of the tonicity agent is about 0.1-1%. In certain embodiments, the concentration of the tonicity agent is about 0.5-3%. In certain embodiments, the concentration of the tonicity agent is about 0.25 wt%. In certain embodiments, the concentration of the tonicity agent is about 0.45 wt%. In certain embodiments, the concentration of the tonicity agent is about 0.9 wt%. In certain embodiments, the concentration of the tonicity agent is about 1.2 wt%. In certain embodiments, the concentration of the tonicity agent is about 2.4 wt%. In certain embodiments, the concentration of the tonicity agent is about 5 wt%.

In some embodiments, the tonicity agent may be present in the composition and/or formulation in one or more of the aforementioned ranges during the formation and/or dilution processes described herein. In certain embodiments, the tonicity agent may be present in the composition and/or formulation in one or more of the aforementioned ranges in the final product.

It is recognized in the art that the ionic strength of a formulation comprising particles can affect the polydispersity of the particles. Polydispersity is a measure of the non-uniformity of particle size in a formulation. The non-uniformity in particle size may be due to the presence of agglomeration and/or differences in individual particle sizes in the formulation. When the particles have essentially the same size, shape, and/or mass, the formulation comprising the particles is considered to be substantially homogeneous or “monodisperse”. Formulations comprising particles of various sizes, shapes, and/or masses are considered to be heterogeneous or “polydisperse”.

The ionic strength of the formulation containing the particles can also affect the colloidal stability of the particles. For example, the relatively high ionic strength of the formulation can lead to coagulation of the particles of the formulation and thus render the formulation unstable. In some embodiments, formulations comprising particles are stabilized by interparticle repulsion forces. For example, the particles are electrically or electrostatically charged. The two charged particles can repel each other and prevent collision and agglomeration. When the repulsion between the particles weakens or becomes attractive, the particles can begin to agglomerate. For example, when the ionic strength of the formulation is increased to a certain level, the charge of the particles (e.g., negative charge) is neutralized by oppositely charged ions (e.g., Na ⁺ ions in solution) present in the formulation. Can be. As a result, the particles can collide and combine with each other to form larger sized aggregates (eg, clusters or flocs). The formed agglomerates of the particles may also differ in size, thus increasing the polydispersity of the formulation. For example, if the ionic strength of the formulation is increased beyond a certain level, a formulation comprising particles of similar size can be a formulation comprising particles of various sizes (eg, due to agglomeration). In the process of agglomeration, the agglomerates may grow in size and eventually settle to the bottom of the container, and the formulation is considered colloidal unstable. Once the particles in the formulation form agglomerates, it is usually difficult to crush the agglomerates into individual particles.

Certain formulations described herein, inter alia, the presence of one or more ionic tonicity agents (e.g., salts such as NaCl) in the formulation at a specific concentration actually reduces or maintains the degree of agglomeration of the particles present in the formulation and/ Or, it exhibits unexpected properties in that it does not significantly increase aggregation. In certain embodiments, the polydispersity of the formulation is relatively constant upon addition of one or more ionic tonicity agents to the formulation, or does not change with appropriate amounts.

For example, in some embodiments, in the presence of the added ionic strength and/or the added ionic strength of the composition and/or formulation remains relatively constant or increases (e.g., during the formation and/or dilution process ) The polydispersity of the composition and/or formulation is relatively constant. In certain embodiments, when the ionic strength increases by 50% or more, the polydispersity is about 200% or less, about 150% or less, about 100% or less, about 75% or less, about 50% or less, about 30% or less, about Increase to less than 20%, less than about 10%, less than about 3%, or less than about 1%. In certain embodiments, when the ionic strength is increased by at least 50%, the polydispersity increases by at least about 1%, at least about 3%, at least about 10%, at least about 30%, or at least about 100%. Combinations of the above-mentioned ranges are possible (for example, an increase in polydispersity of 50% or less and 1% or more). Other ranges are also possible.

The ionic strength of the formulations described herein can be controlled (eg, increased) by various means, such as adding one or more ionic tonicity agents (eg, salts such as NaCl) to the formulation. In certain embodiments, the ionic strength of the formulations described herein is at least about 0.0005 M, at least about 0.001 M, at least about 0.003 M, at least about 0.01 M, at least about 0.03 M, at least about 0.1 M, at least about 0.3 M, about 1 M or more, about 3 M or more, or about 10 M or more. In certain embodiments, the ionic strength of the formulations described herein is about 10 M or less, about 3 M or less, about 1 M or less, about 0.3 M or less, about 0.1 M or less, about 0.03 M or less, about 0.01 M or less, about 0.003 M or less, about 0.001 M or less, or about 0.0005 M or less. Combinations of the above-mentioned ranges are possible (eg, ionic strength of about 0.01 M or more and about 1 M or less). Other ranges are also possible. In certain embodiments, the ionic strength of the formulations described herein is about 0.1 M. In certain embodiments, the ionic strength of the formulations described herein is about 0.15 M. In certain embodiments, the ionic strength of the formulations described herein is about 0.3 M.

In certain embodiments, the polydispersity of the formulation does not change upon addition of one or more ionic tonicity agents to the formulation. In certain embodiments, the polydispersity does not increase significantly upon addition of one or more ionic tonicity agents to the formulation. In certain embodiments, the polydispersity increases to the levels described herein upon addition of one or more ionic tonicity agents to the formulation.

The polydispersity of the formulations described herein can be measured by the polydispersity index (PDI). PDI is used to describe the width of the particle size distribution and is often calculated from cumulants analysis of dynamic light scattering (DLS) measured intensity autocorrelation functions. The calculation of these parameters is specified in the standards ISO 13321: 1996 E and ISO 22412: 2008. PDI is dimensionless and, as measured by DLS, values less than 0.05 indicate highly monodisperse samples, while values greater than 0.7 are normalized to indicate a very wide size distribution. In certain embodiments, the PDI of the formulations and/or compositions described herein is about 1 or less, about 0.9 or less, about 0.8 or less, about 0.7 or less, about 0.6 or less, about 0.5 or less, about 0.4 or less, about 0.3 or less, about 0.2 Or less, about 0.15 or less, about 0.1 or less, about 0.05 or less, about 0.01 or less, or about 0.005 or less. In certain embodiments, the PDI of the formulations and/or compositions described herein is at least about 0.005, at least about 0.01, at least about 0.05, at least about 0.1, at least about 0.15, at least about 0.2, at least about 0.3, at least about 0.4, about 0.5 Or more, about 0.6 or more, about 0.7 or more, about 0.8 or more, about 0.9 or more, or about 1 or more. Combinations of the aforementioned ranges are possible (eg, PDI of about 0.1 or greater and about 0.5 or less). Other ranges are also possible. In certain embodiments, the formulation has a PDI of about 0.1. In certain embodiments, the formulation has a PDI of about 0.15. In certain embodiments, the formulation has a PDI of about 0.2.

In certain embodiments, the compositions and/or formulations described herein may be highly dispersible and do not tend to form aggregates. Even when the particles form agglomerates, the agglomerates can easily be broken down into individual particles without strictly stirring the composition and/or formulation.

In general, it is preferred that the formulation is sterile prior to or upon administration to a subject. Sterile preparations are generally essentially free of pathogenic microorganisms, such as bacteria, microbes, fungi, viruses, spores, yeast, fungi, etc., associated with infection. In some embodiments, compositions and/or formulations comprising the coated particles described herein can be subjected to aseptic processes and/or other sterilization processes. Aseptic processes typically involve combining processes such as heating, gamma irradiation, ethylene oxide, or filtration to sterilize the formulation, the components of the final formulation, and/or the container closure of the drug product, and in a sterile environment. do. In some cases, aseptic process is preferred. In other embodiments, terminal sterilization is preferred.

Examples of other sterilization methods include radiation sterilization (eg, gamma, electron, or x-ray radiation), heat sterilization, sterile filtration, and ethylene oxide sterilization. The terms "radiation" and "irradiation" are used interchangeably herein. Unlike other sterilization methods, radiation sterilization has the advantage of high permeability and immediate effect, without the need to control temperature, pressure, vacuum, or humidity in some cases. In certain embodiments, the radiation used to sterilize the coated particles described herein is gamma radiation. Gamma radiation can be applied in an amount sufficient to kill most or substantially all of the bacteria in or on the coated particles. The temperature and rate of radiation of the coated particles described herein can be relatively constant over the entire gamma radiation period. Gamma irradiation can be carried out at any suitable temperature (eg, ambient temperature, about 40° C., about 30 to about 50° C.). Unless otherwise specified, measurements of gamma irradiation described herein refer to those performed at about 40°C.

In embodiments using a sterilization process, the process comprises: (1) not significantly changing the particle size of the coated particles described herein; (2) not significantly altering the integrity of the active ingredient (eg drug) of the coated particles described herein; And (3) it may be desirable not to generate impurities of an unacceptable concentration during or after the process. In certain embodiments, impurities generated during or after processing are a degradant of the active ingredient of the coated particles described herein.

The compositions and/or formulations described herein can have any suitable pH value. The term “pH”, unless otherwise provided, refers to the pH measured at ambient temperature (eg, about 20° C., about 23° C., or about 25° C.). The composition and/or agent has, for example, an acidic pH, a neutral pH, or a basic pH, and can be determined, for example, by where the composition and/or agent is delivered into the body. In certain embodiments, the composition and/or formulation has a physiological pH. In certain embodiments, the pH value of the composition and/or formulation is at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 6.2, at least about 6.4, at least about 6.6, About 6.8 or more, about 7 or more, about 7.2 or more, about 7.4 or more, about 7.6 or more, about 7.8 or more, about 8 or more, about 8.2 or more, about 8.4 or more, about 8.6 or more, about 8.8 or more, about 9 or more, about 10 Or more, about 11 or more, or about 12 or more. In certain embodiments, the pH value of the composition and/or formulation is about 12 or less, about 11 or less, about 10 or less, about 9 or less, about 8.8 or less, about 8.6 or less, about 8.4 or less, about 8.2 or less, about 8 or less, About 7.8 or less, about 7.6 or less, about 7.4 or less, about 7.2 or less, about 7 or less, about 6.8 or less, about 6.6 or less, about 6.4 or less, about 6.2 or less, about 6 or less, about 5 or less, about 4 or less, about 3 Or less, about 2 or less, or about 1 or less. Combinations of the above-mentioned ranges are possible (eg, a pH value of about 5 or more and about 8.2 or less). Other ranges are also possible. In certain embodiments, the compositions and/or formulations described herein have a pH value of about 5 or more and about 8 or less.

In one set of embodiments, a composition comprising a plurality of coated particles is provided. The coated particles include core particles comprising a medicament or a salt thereof, and a coating comprising a surface modifier surrounding the core particles. The core particles comprising the medicament or a salt thereof may be, for example, solid and at any time point throughout the pH range of about 1 mg/mL or less at 25° C. (e.g., about 0.1 mg/mL or less at 25° C. ). However, other cores (eg, polymer cores) can be used as described herein. Surface modifiers are or include synthetic polymers having pendant hydroxyl groups on the backbone of the polymer. The polymer has a molecular weight of about 1 kDa or more and about 1000 kDa or less (e.g., about 200 kDa or less), and about 30% or more hydrolysis (e.g., about 70% hydrolysis or more) about 98% or less hydrolysis (E.g., less than about 95% hydrolyzed). The synthetic polymer can be, for example, poly(vinyl alcohol), partially hydrolyzed poly(vinyl acetate) or a copolymer of vinyl alcohol and vinyl acetate. (In certain embodiments, the polymer is less than about 98% hydrolyzed and has a molecular weight of less than about 75 kDa, or less than about 95% hydrolyzed PVA). The surface modifier can be adsorbed on the surface of the core particles. In certain cases, the surface modifier is present on the surface of the core particle at a density of at least about 0.001 molecules per nanometer square (eg, at least about 0.01 molecules per nanometer square). The coated particles have a relative velocity of greater than 0.5 in the mucus. The coated particles may have an average size of about 20 nm or more and about 1 μm or less (eg, about 500 nm or less). The thickness of the coating can be, for example, about 100 nm or less (eg, about 50 nm or less, about 30 nm or less, about 10 nm or less). The composition may be a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, additives, and/or diluents. The total amount of surface modifier present in the composition is, for example, from about 0.001% to about 5% by weight (e.g., from about 0.01% to about 5% by weight, or from about 0.1% to about 5% by weight). I can. In some embodiments, the composition has a PDI of about 0.1 or greater and about 0.5 or less (eg, about 0.3 or less or about 0.2 or less). Optionally, in some embodiments, the plurality of coated particles are in solution (eg, an aqueous solution) with one or more glass surface modifiers in the composition. The glass surface modifying agent(s) in solution and the surface modifying agent on the surface of the particles may be the same (or different) surface modifying agent(s) and may equilibrate with each other in the composition. Methods of use and/or delivery of such compositions (eg, to mucus or mucous membranes) to a patient or subject are also provided.

In one set of embodiments, a composition comprising a plurality of coated particles is provided. The coated particles include core particles comprising a drug or a salt thereof, and a coating surrounding the core particles. The core particles comprising the medicament or a salt thereof may be, for example, solid and at any time point throughout the pH range of about 1 mg/mL or less at 25° C. (e.g., about 0.1 mg/mL or less at 25° C. ). However, other cores (eg, polymer cores) can be used as described herein. The surface modifier may be or include poly(vinyl alcohol), wherein the poly(vinyl alcohol) is PVA 13K87, PVA 31K98, PVA 31K87, PVA 9K80, PVA 2K75, PVA 57K87, PVA 85K87, PVA105K80, PVA130K87, where " The numbers before and after K" are each selected from the molecular weight of the PVA in kDa and the degree of hydrolysis in %), and combinations thereof. The surface modifier can be adsorbed on the surface of the core particles. In certain cases, the surface modifier is present on the surface of the core particle at a density of at least about 0.001 molecules per nanometer square (eg, at least about 0.01 molecules per nanometer square). The coated particles have a relative velocity of greater than 0.5 in the mucus. The coated particles may have an average size of about 20 nm or more and about 1 μm or less (eg, about 500 nm or less). The thickness of the coating can be, for example, about 100 nm or less (eg, about 50 nm or less, about 30 nm or less, about 10 nm or less). The composition may be a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, additives, and/or diluents. The total amount of surface modifier present in the composition is, for example, from about 0.001% to about 5% by weight (e.g., from about 0.01% to about 5% by weight, or from about 0.1% to about 5% by weight). I can. In some embodiments, the composition has a PDI of about 0.1 or greater and about 0.5 or less (eg, about 0.3 or less or about 0.2 or less). Optionally, in some embodiments, the plurality of coated particles are in solution (eg, an aqueous solution) with one or more glass surface modifiers in the composition. The glass surface modifying agent(s) in solution and the surface modifying agent on the surface of the particles may be the same (or different) surface modifying agent(s) and may equilibrate with each other in the composition. Methods of use and/or delivery of such compositions (eg, to mucus or mucous membranes) to a patient or subject are also provided.

These and other aspects of the invention will be further recognized upon consideration of the following examples, which are intended to illustrate certain specific embodiments of the invention, but limit their scope, as defined by the claims. I don't want to.

Example

Example One

The following describes a non-limiting example of a method of forming mucus-penetrating particles from pre-assembled polymer particles by physical adsorption of certain poly(vinyl alcohol) polymers (PVA). Carboxylated polystyrene nanoparticles (PSCOO) were used as pre-assembled particles/core particles with strong mucoadhesive behavior fixed. PVA acted as a surface modifier to form a coating around the core particles. PVA of various molecular weights (MW) and degree of hydrolysis were evaluated to determine the effectiveness of the coated particles in penetrating mucus.

PSCOO particles are incubated in an aqueous solution in the presence of various PVA polymers to minimize particle interaction with mucus constituents and cause rapid particle penetration in mucus. (Non-covalently) coating was determined. In these experiments, PVA acted as a coating around the core particles, and the resulting particles were tested for their mobility in mucus, but in other embodiments, PVA is another surface modifier that can increase the mobility of particles in mucus. Can be replaced with. The PVAs tested ranged from 2 kDa to 130 kDa with an average molecular weight and an average degree of hydrolysis of 75% to 99+%. The PVAs tested are listed in Table 1 shown above.

The particle modification process was as follows: 200 nm carboxylation-modified red fluorescent polystyrene nanoparticles (PSCOO) were purchased from Invitrogen. PSCOO particles (0.4-0.5% wt) were incubated in an aqueous PVA solution (0.4-0.5% wt) for at least 1 hour at room temperature.

The mobility and distribution of the modified nanoparticles in human cervical mucus (CVM) was characterized using fluorescence microscopy and multiple particle tracking software. In a typical experiment, ≤0.5 μL of incubated nanosuspension (˜10× dilution with a 0.5% wt aqueous solution of the corresponding PVA) was added to 20 μl of fresh CVM along with the control. Conventional nanoparticles (200 nm blue fluorescent carboxylate-modified polystyrene microspheres, obtained from Invitrogen) were used as negative controls to confirm the barrier properties of CVM samples. Yellow-green fluorescent polystyrene nanoparticles covalently coated with PEG 2 kDa were used as a positive control with fixed MPP behavior. Using a fluorescence microscope equipped with a CCD camera, each type of particle, i.e., a sample (observed through a Texas Red filter set), a negative control (observed through a DAPI filter set), and a positive control (FITC filter) A 15 s movie was captured at a temporal resolution of 66.7 ms (15 frames/s) under 100x magnification from several regions within each sample for observation through the set). Then, using advanced image processing software, individual trajectories of the multiple particles were measured over a time scale of 3.335 s (50 frames) or more. The resulting transport data are here in the form of the orbital-average velocity V _average , i.e. the velocity of the individual particles averaged over its orbit, and the ensemble-average velocity <V _mean >, i.e. the V _average averaged over the ensemble of particles. Presented. In order to allow easy comparison between different samples and to define the velocity data for the natural variability of the penetrability of the CVM samples, the ensemble-averaged (absolute) velocity is then converted to the relative sample rate <V according to the equation shown in Equation 1 Switched to _average > _relative . The negative control was constrained in all tested CVM samples, while the positive control was confirmed to be mobile (Table 3), as evidenced by the difference in <V _mean > for the positive and negative control according to multi-particle tracking (Table 3). .

It was found that nanoparticles incubated in the presence of certain (but, interestingly, not all) PVAs, were transported through the CVM at the same rate or at about the same rate as the positive control. Specifically, the particles stabilized with PVA2K75, PVA9K80, PVA13K87, PVA31K87, PVA57K86, PVA85K87, PVA105K80, and PVA130K87 significantly exceed the <V _mean > of the negative control, and within the experimental error <V _mean , which is indistinguishable from that of the positive control group. >. The results are shown in Table 3 and Fig. 2A. For these samples, the <V _mean > _relative value exceeded 0.5, as shown in Figure 2B.

On the other hand, nanoparticles incubated with PVA95K95, PVA13K98, PVA31K98, and PVA85K99 were mostly or completely immobilized as evidenced by each <V _mean > _relative value of 0.1 or less (Table 3 and Figure 2B).

In order to confirm the characteristics of PVA that allows the particles to penetrate mucous, the <V _average > _relative of the nanoparticles prepared by incubation with various PVAs was mapped to the MW and degree of hydrolysis of the PVA used (Fig. 3). At least, it was concluded that such a PVA with a degree of hydrolysis of less than 95% made the nanocrystals muco-penetrating. Without wishing to be bound by any theory, the non-hydrolyzed (vinyl acetate) units of the PVA are sufficient (e.g., greater than 5% in some embodiments) of the surface of the core particles if the content of these segments in the PVA is sufficient. Can provide effective hydrophobic association; On the one hand, it is believed that the hydrophilic (vinyl alcohol) units of PVA present on the surface of the coated particles make them hydrophilic and can shield the coated particles from adhesion interactions with mucus.

To further confirm the ability of specific PVA grades to convert mucoadhesive particles into muco-penetrating particles by physical adsorption, PSCOO nanoparticles incubated with various PVAs were tested using a bulk transport assay. In this method, 20 μL of CVM was collected in a capillary tube and one end was sealed with clay. The open end of the capillary was then immersed in an aqueous suspension of 20 μL of particles (which is 0.5% w/v drug). After the desired time, typically 18 hours, the capillaries were removed from the suspension and the exterior was wiped clean. The capillary tube containing the mucus sample was placed in the ultracentrifuge tube. The extraction medium was added to the tube and incubated for 1 hour while mixing, which was to remove the mucus from the capillary and extract the drug from the mucus. The sample was then spun to remove mucin and other insoluble components. Then, the amount of drug in the extracted sample could be quantified using HPLC. The results of these experiments are in close agreement with those of microscopy, demonstrating a clear differentiation in transport between positive (mucus-penetrating particles) and negative controls (conventional particles). The bulk transport results for PSCOO nanoparticles incubated with various PVAs are shown in FIG. 4. These results confirm the findings for microscopy/particle tracking using PSCOO nanoparticles incubated with various PVAs and demonstrate that incubating the nanoparticles with partially hydrolyzed PVA improves mucus penetration.

Example 2

The following describes a non-limiting example of a method of forming mucus-penetrating particles by an emulsification process in the presence of certain poly(vinyl alcohol) polymers (PVA). Polylactide (PLA), a biodegradable pharmaceutical-related polymer, was used as a material for forming core particles through an oil-in-water emulsification process. PVA served as a surface modifier and emulsion stabilizer to form a coating around the prepared core particles. PVA of various molecular weights (MW) and degree of hydrolysis were evaluated to determine the effectiveness of the formed particles in penetrating mucus.

PLA solution in dichloromethane can be physically (non-covalently) coated on the surface of specific PVA-generated nanoparticles with a coating that will cause rapid particle penetration in mucus by emulsifying in aqueous solution in the presence of various PVA polymers. It was decided. In these experiments, PVA served as a surfactant, upon solidification, forming a stabilizing coating around droplets of the emulsified organic phase that formed core particles. The resulting particles were tested for their mobility in mucus, but in other embodiments, PVA can be replaced with other surface modifiers that can increase the mobility of the particles in mucus. The PVAs tested ranged from 2 kDa to 130 kDa with an average molecular weight and an average degree of hydrolysis of 75% to 99+%. The PVAs tested are listed in Table 1 shown above.

The emulsion-solvent evaporation process was as follows: a solution of approximately 0.5 mL, 20-40 mg/mL PLA (polylactide grade 100DL7A, purchased from Surmodics) in dichloromethane was approximately 4 mL by sonication. Emulsification in aqueous PVA solution (0.5-2% wt) gave a stable emulsion with a target number-average particle size of <500 nm. The obtained emulsion was immediately subjected to thorough rotary evaporation under reduced pressure at room temperature to remove the organic solvent. The resulting suspension was filtered through a 1 micron glass fiber filter to remove any aggregates. Table 4 lists the particle size characteristics of the nanosuspensions obtained by this emulsification procedure using various PVAs. In all cases, fluorescent organic dye Nile Red was added to the emulsified organic phase to fluorescently label the resulting particles.

The mobility and distribution of the prepared nanoparticles in human cervical mucus (CVM) was characterized using fluorescence microscopy and multiple particle tracking software. In a typical experiment, ≤0.5 uL of nanosuspension (if necessary, diluted to a PVA concentration of -0.5 wt%) was added to 20 μl of fresh CVM along with the control. Conventional nanoparticles (200 nm blue fluorescent carboxylate-modified polystyrene microspheres, obtained from Invitrogen) were used as negative controls to confirm the blocking properties of CVM samples. Yellow-green fluorescent polystyrene nanoparticles covalently coated with PEG 2 kDa were used as a positive control with fixed MPP behavior. Using a fluorescence microscope equipped with a CCD camera, each type of particle, i.e., a sample (observed through a Texas Red filter set due to encapsulated Nile Red), a negative control (observed through a DAPI filter set), and a positive control ( A 15 s movie was captured at a temporal resolution of 66.7 ms (15 frames/s) under 100x magnification from several regions within each sample for observation through a FITC filter set). Then, using advanced image processing software, individual trajectories of the multiple particles were measured over a time scale of 3.335 s (50 frames) or more. The resulting transport data are here in the form of the orbital-average velocity V _average , i.e. the velocity of the individual particles averaged over its orbit, and the ensemble-average velocity <V _mean >, i.e. the V _average averaged over the ensemble of particles. Presented. In order to allow easy comparison between different samples and to define the velocity data for the natural variability of the penetrability of the CVM samples, the ensemble-averaged (absolute) velocity is then converted to the relative sample rate <V according to the equation shown in Equation 1 Switched to _average > _relative . The negative control was constrained in all tested CVM samples, while the positive control was confirmed to be mobile (Table 5), as evidenced by the difference in <V _mean > for the positive and negative control according to multi-particle tracking (Table 5). .

It has been found that nanoparticles prepared in the presence of certain (but, interestingly, not all) PVAs, were transported through the CVM at the same rate or at about the same rate as the positive control. Specifically, particles stabilized with PVA2K75, PVA9K80, PVA13K87, PVA31K87, PVA85K87, PVA105K80, and PVA130K87 significantly exceed the <V _mean > of the negative control group and are positive within the experimental error, as shown in Table 5 and FIG. 5A. The <V _mean > was shown, which was indistinguishable from that of the control group. For these samples, the <V _mean > _relative value exceeded 0.5, as shown in Figure 5B.

On the other hand, pyrene nanoparticles obtained using PVA95K95, PVA13K98, PVA31K98, and PVA85K99 were mostly or completely immobilized as evidenced by each <V _mean > _relative value of 0.4 or less (Table 5 and Figure 5B). Became. In order to confirm the characteristics of PVA that allows the particles to penetrate mucus, the <V _average > _relative of the nanoparticles prepared using various PVAs was mapped to the MW and degree of hydrolysis of the PVA used (FIG. 6). At least, it was concluded that such a PVA with a degree of hydrolysis of less than 95% made the nanocrystals muco-penetrating. Without wishing to be bound by any theory, the non-hydrolyzed (vinyl acetate) units of the PVA are sufficient (e.g., greater than 5% in some embodiments) of the surface of the core particles if the content of these segments in the PVA is sufficient. Can provide effective hydrophobic association; On the other hand, it is believed that the hydrophilic (vinyl alcohol) units of PVA present on the surface of the coated particles make them hydrophilic and can shield the coated particles from adhesive interactions with mucus.

Example 3

The following describes a non-limiting example of a method of forming mucus-penetrating non-polymeric solid particles by nanomilling in the presence of certain poly(vinyl alcohol) polymers (PVA). Pyrene, a hydrophobic model compound, was used as core particles processed by nanomilling. PVA served as a surface modifier to form a coating around the core particles and as a nanomilling aid to accelerate the particle size reduction of the core particles. PVA of varying molecular weight (MW) and degree of hydrolysis were evaluated to determine the effectiveness of nanomilled particles in penetrating mucus.

By nano-milling pyrene in an aqueous dispersion in the presence of various PVAs, whether a specific MW and PVA of a degree of hydrolysis can help 1) reduce the particle size to several hundred nanometers and 2) minimize particle interaction with mucus components. It was determined whether it was possible to physically (non-covalently) coat the surface of the resulting nanoparticles with a mucosal inert coating that would prevent mucus adhesion. In these experiments, PVA served as a coating around the core particles, and the resulting particles were tested for their mobility in mucus. The PVAs tested ranged from 2 kDa to 130 kDa with an average molecular weight and an average degree of hydrolysis of 75% to 99+%. The PVAs tested are listed in Table 1 shown above. Various other polymers listed in Table 6, including pharmaceutically relevant excipients such as polyvinylpyrrolidone (Kollidon), hydroxypropyl methylcellulose (Methocel), Tween, Span, etc. , Oligomers, and small molecules were tested in a similar manner.

The aqueous dispersion containing pyrene and one of the stabilizers/surface modifiers listed above was stirred with the milling medium until the particle size (as measured by dynamic light scattering) was reduced to less than 500 nm. Table 7 lists the particle size characteristics of the pyrene particles obtained by nanomilling in the presence of various stabilizers/surface modifiers. When Span 20, Span 80, or octyl glucoside was used as a stabilizer/surface modifier, a stable nanosuspension could not be obtained. Therefore, these stabilizers/surface modifiers were excluded from further investigation due to their inability to effectively assist in particle size reduction.

The mobility and distribution of the prepared pyrene nanoparticles in human cervical mucus (CVM) was characterized using fluorescence microscopy and multi-particle tracking software. In a typical experiment, ≤0.5 uL of nanosuspension (if necessary, diluted to a concentration of ˜1% surfactant) was added to 20 μl of fresh CVM along with the control. Conventional nanoparticles (200 nm yellow-green fluorescent carboxylate-modified polystyrene microspheres, obtained from Invitrogen) were used as negative controls to confirm the blocking properties of CVM samples. Red fluorescent polystyrene nanoparticles covalently coated with PEG 5 kDa were used as a positive control with fixed MPP behavior. Using a fluorescence microscope equipped with a CCD camera, each type of particle, i.e., a sample (pyrene), a negative control, and a positive control (natural blue fluorescence of pyrene, which allowed observation of pyrene nanoparticles separately from the control) was tested. A 15 s movie was captured at a temporal resolution of 66.7 ms (15 frames/s) under 100x magnification from several regions within each sample for. Then, using advanced image processing software, individual trajectories of the multiple particles were measured over a time scale of 3.335 s (50 frames) or more. The resulting transport data are here in the form of the orbital-average velocity V _average , i.e. the velocity of the individual particles averaged over its orbit, and the ensemble-average velocity <V _mean >, i.e. the V _average averaged over the ensemble of particles. Presented. In order to allow easy comparison between different samples and to define the velocity data for the natural variability of the penetrability of the CVM samples, the ensemble-averaged (absolute) velocity is then converted to the relative sample rate <V according to the equation shown in Equation 1 Switched to _average > _relative .

Before quantifying the mobility of the pyrene particles, their spatial distribution in the mucus sample was visually evaluated. The pyrene/methocell nanosuspensions did not achieve a uniform distribution in CVM and were found to aggregate strongly into domains much larger than the mucous mesh size (data not shown). This agglomeration is a sign of mucosal adhesion behavior and effectively prevents mucus penetration. Thus, further quantitative analysis of particle mobility was considered unnecessary. Similar to the positive control, all other tested pyrene/surface modifier systems achieved a fairly uniform distribution among the CVM. The negative control was constrained in all tested CVM samples, while the positive control was confirmed to be mobile, as evidenced by the difference in <V _mean > for the positive and negative controls by multi-particle tracking (Table 8). ).

It was found that the nanoparticles obtained in the presence of certain (but, interestingly, not all) PVA, were transported through the CVM at the same rate or at about the same rate as the positive control. Specifically, pyrene nanoparticles stabilized with PVA2K75, PVA9K80, PVA13K87, PVA31K87, PVA85K87, and PVA130K87 significantly exceed the <V _mean > of the negative control group and are positive within the experimental error, as shown in Table 8 and FIG. The <V _mean > was shown, which was indistinguishable from that of the control group. For these samples, the <V _mean > _relative value exceeded 0.5, as shown in Figure 7B.

On the other hand, pyrene nanoparticles obtained using other stabilizers/surface modifiers, including PVA95K95, PVA13K98, PVA31K98, and PVA85K99, were largely as evidenced by each <V _average > _relative value of 0.5 or less. Alternatively, it was completely immobilized and was 0.4 or less when most of the stabilizers/surface modifiers were used (Table 8 and FIG. 7B). In addition, Figs. 8A-8F are histograms showing the distribution of V _averages within the ensemble of particles. These histograms show the muco-diffusion (muco-diffusion) of samples stabilized with PVA2K75 and PVA9K80 in contrast to the mucoadhesion behavior of samples stabilized with PVA31K98, PVA85K99, Collidone 25, and Colicoat IR (selected as a representative mucoadhesive sample). diffusive) behavior (a similar histogram was obtained for samples stabilized with PVA13K87, PVA31K87, PVA85K87, and PVA130K87, but not shown here).

In order to confirm the characteristics of PVA that allows the pyrene nanocrystals to penetrate mucous, the <V _average > _relative of the pyrene nanocrystals stabilized using various PVAs was mapped to the MW and the degree of hydrolysis of the PVA used (Fig. 9). . At least, it was concluded that such a PVA with a degree of hydrolysis of less than 95% made the nanocrystals muco-penetrating. Without wishing to be bound by any theory, the non-hydrolyzed (vinyl acetate) segments of PVA may be found in the surface of the core particles if the content of these segments in the PVA is sufficient (e.g., greater than 5% in some embodiments). Can provide effective hydrophobic association; Meanwhile, it is believed that the hydrophilic (vinyl alcohol) segments of PVA present on the surface of the coated particles make them hydrophilic and can shield the coated particles from adhesion interactions with mucus.

Example 4

These non-limiting embodiment is compared with the Pluronic ^® F 127, Tween 80 ^®, or LE MPP containing PVA is Rotterdam Max ^® as surface modifiers demonstrates that shows the improved exposure of LE in rabbits.

Two additional surface modifiers, Tween 80 ^® and polyvinyl alcohol (PVA) were studied to demonstrate that LE MPP's ability to enhance the exposure of LE is not limited to the inclusion of F127 as a surface modifier in LE MPP. I did. Tween 80 ^® is an FDA approved surface modifier consisting of a PEGylated sorbitan and an alkyl tail forming a head group. Tween 80 ^® differs from the range of other surface modifiers (eg F127 and PVA), inter alia, in that it is oligomeric and thus has a significantly lower molecular weight. PVA is a FDA approved polymer prepared, for example by partially hydrolyzing polyvinyl acetate to create a random copolymer of polyvinyl acetate and polyvinyl alcohol. PVA is, among others, which is different from the range of other surface modification in that they do not contain any claim PEG (e. G., F127 and Tween 80 ^®). It was shown earlier than expected that some PVAs allowed mucus penetration, while others did not. This differentiated mucus penetration behavior can be controlled by the molecular weight and degree of hydrolysis of PVA. Based on the results from these experiments, PVA with a molecular weight of about 2 kDa and about 75% hydrolyzed was chosen for the study of the mucus penetrating properties of LE MPP.

To form the LE MPP, the milling procedure was used as described herein. In one set of embodiments, an aqueous dispersion containing one surface modifier selected from LE and F127, Tween 80 ^® , and PVA (2 kDa, 75% hydrolyzed) is prepared as the particle size is determined by dynamic light scattering. Milled with grinding media until reduced to less than about 300 nm. Mucus mobility was characterized in mucus of the human cervix based on the previously described characterization method. All three LE MPPs (ie, LE-F127, LE-Twin80, and LE-PVA) exhibited mucus penetrating properties (FIG. 10).

In vivo, due to the single topical infusion of one of the three LE MPPs in New Zealand white rabbits, LE levels in the rabbit's cornea were significantly higher than the LE levels from Rotemax ^{® at a} similarly administered dose. Resulted (Figure 10). These results demonstrate that MPPs, compositions, and/or formulations containing the drug enhance the exposure of the drug based on the mucus penetrating properties of the particles.

Example 5

The following non-limiting examples demonstrate the evaluation of different surface modifiers (e.g., poly(vinyl alcohol) (PVA)) in preparing mucus-penetrating particles comprising loteprednol etabonate (LE) as the core material. Write.

In this example, LE, a relatively hydrophobic agent, was milled as an aqueous suspension with milling media in the presence of various types of PVA as surface modifiers. The characteristics of the PVA tested, including molecular weight (MW) and degree of hydrolysis, are listed in Table 9. The milling process was carried out until the LE particles became uniformly small (i.e., Z-average diameter (D) ≤500 nm and polydispersity index (PDI) <0.20 as measured by dynamic light scattering (DLS)). In addition, the resulting particle size and polydispersity index of LE nanoparticles as measured by DLS are listed in Table 9.

The mucus-penetrating ability of LE nanoparticles from the prepared nanosuspensions was characterized in cervical mucus (CVM) (as described in Example 2) restored by high resolution cancer-based microscopy. In a typical sample formulation, 0.5 μL of nanosuspension was placed on a microscope slide at a nanoparticle concentration of 20 μL of rehydrated CVM and 5% w/v LE. In the case of LE milled in DPPC or DPPC+2PEG-PE, 1 μL of nanosuspension was used at a concentration of 1% w/v LE. Video at a 15 s length and a temporal resolution of 66.7 ms (15 frames/s) was acquired for randomized areas within mucus samples under dark field microscopy at high magnification (100x). Settled positive control (LE nanoparticles milled with Pluronic ^® F127, “mobility” or “muco-penetrating”) and negative control (LE nanoparticles milled with SDS, “not mobile” or “not mucus-penetrating” ) To determine the mobility of LE nanoparticles in the video visually by comparing the degree of movement of the particles in the recorded video (i.e., the speed of the entire particle and the amount of particles seen as mobility), and "mobility" and "mobility, respectively. Classified as "not".

In order to confirm the characteristics of PVA that allows LE nanocrystals to penetrate mucous, the mobility for LE nanoparticles obtained by milling using various PVAs was mapped to the MW and degree of hydrolysis of the PVA used (FIG. 11). . It was observed that such PVAs with both hydrolysis degree >95% and molecular weight >31 kDa did not produce stable nanocrystals or mucus-penetrating ones. Without wishing to be bound by any theory, the non-hydrolyzed (vinyl acetate) segments of PVA may be found in the surface of the core particles if the content of these segments in the PVA is sufficient (e.g., 2% or more in some embodiments). Can provide effective hydrophobic association; On the other hand, it is believed that the hydrophilic (vinyl alcohol) segment of PVA present on the surface of the coated particles makes the coated particles hydrophilic and can shield the coated particles from adhesion interactions with mucus. The results suggest that the following surface modifiers render LE nanoparticles mucus-penetrating: PVA 2K75, PVA 13K87, PVA 31K87, PVA 31K98, PVA 85K87, and PVA 130K87.

Other embodiments

While some embodiments of the present invention have been described and illustrated herein, those skilled in the art will readily envision various other means and/or structures to perform a function and/or obtain a result and/or one or more of the advantages described herein. , Such changes and/or modifications are each considered to be within the scope of the present invention. More generally, one skilled in the art is meant to mean that all parameters, dimensions, materials, and configurations described herein are exemplary, and actual parameters, dimensions, materials, and/or configurations are specific to the specific application or applications for which the teachings of the present invention are used. It will be readily appreciated that it will be determined by One of skill in the art, using mere routine experimentation, will recognize, or will be able to ascertain, many equivalents to the specific embodiments of the invention described herein. Accordingly, it is to be understood that the foregoing embodiments are presented by way of example only, and within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise as specifically described and claimed. The invention is directed to each individual feature, system, article, material, kit, and/or method described herein. In addition, any combination of two or more of these features, systems, articles, materials, kits, and/or methods may be subject to the present invention unless such features, systems, articles, materials, kits, and/or methods are inconsistent with each other. It is included within the scope of the invention.

As used herein in the specification and claims, the indefinite articles “a” and “an” are to be understood to mean “at least one” unless expressly stated otherwise.

As used herein in the specification and claims, the phrase “and/or” means “one or both” of the elements so conjoined, ie, in some cases in combination and in other cases separately. It should be understood to mean the element. Unless expressly stated otherwise, other elements may optionally be present in addition to the elements specifically identified by the "and/or" clause, whether related or not related to those elements specifically identified. Thus, as a non-limiting example, reference to “A and/or B” when used in conjunction with an open-ended language such as “comprising”, in one embodiment, is A without B (optionally B In addition to the elements); In another embodiment, B without A (optionally including elements other than A); In another embodiment, both A and B (optionally including other elements) may refer to and the like.

As used herein in the specification and claims, "or" is to be understood to have the same meaning as "and/or" as defined above. For example, when separating an item from a list, "or" or "and/or" are all-inclusive, ie include at least one of the number or list of elements, but also more than one, and optionally, additional It should be construed as including items not listed. The only terms expressly specified otherwise, such as "only one of" or "exactly one of" or, when used in the claims, "consisting of" shall refer to the inclusion of exactly one element of the list or number of elements. will be. In general, the term “or” as used herein is an exclusive alternative if it is preceded by a term of exclusivity, such as “either”, “one of”, “only one of”, or “exactly one of”. (Ie, "one or the other, but not both") should only be understood. "Consisting essentially of", when used in the claims, should have its ordinary meaning as used in the field of patent law.

As used herein in the specification and claims, the phrase “at least one” means at least one element selected from any one or more of the elements in the list of elements, with respect to the list of one or more elements, but an element It is to be understood that it does not necessarily include at least one of each and every element specifically listed in the list of elements and does not exclude any combination of elements from the list of elements. This definition also allows an element to be present in addition to the specifically identified element in the list of elements to which the phrase “at least one” refers, whether related to or not related to that element specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B”, or, equivalently “at least one of A and/or B”) is one embodiment At least one A (and optionally including elements other than B), wherein no B is present and optionally comprises more than one; In another embodiment, at least one B (and optionally including an element other than A), without any A present, optionally comprising more than one; In another embodiment, it may refer to at least one A, optionally comprising more than one, and at least one B (and optionally including other elements), optionally comprising more than one, and the like.

In the claims, as well as in the specification, all transitional phrases, such as "comprising", "including", "jinin", "having", "including", "involving"","holding," and the like are to be understood as meaning open-ended, ie, including but not limited to. "Consisting only of" and "consisting essentially of" must be the closed or semi-closed transitional phrases specified in section 2111.03 of the manual of the US Patent and Trademark Office's patent examination procedures, respectively.

Claims (47)

Each of the coated nanoparticles
Core particles comprising a drug or a salt thereof; And
Coating comprising a surface modifier surrounding the core particles
Including, where
The drug constitutes at least 80 wt% of the core particles,
The surface modifier is poly(vinyl alcohol) having a molecular weight of 2 kDa or more and 130 kDa or less, wherein the poly(vinyl alcohol) has a degree of hydrolysis of 75% or more and less than 95%,
The coated nanoparticles have a relative velocity <V _average > _relative in mucus of greater than 0.5,
here

Is
Where <V _mean > is the ensemble average trajectory-mean velocity, V _mean is the velocity of the individual particles _averaged over their trajectory, the sample is the particle of interest, and the negative control is 200 nm carboxylate. Misfired polystyrene particles, positive control is 200 nm polystyrene particles densely PEGylated with 2 kDa-5 kDa poly(ethylene glycol),
A composition comprising a plurality of coated nanoparticles. The composition of claim 1 wherein the surface modifier is covalently attached to the core particles. The composition of claim 1 wherein the surface modifier is non-covalently adsorbed to the core particles. The composition of claim 1, wherein the coating is a coating of poly(vinyl alcohol). The composition of claim 1 or 4, wherein the core particles are free of polymeric components. The composition according to claim 1 or 4, wherein the poly(vinyl alcohol) has a molecular weight of 5 kDa or more, 10 kDa or more, 20 kDa or more, 50 kDa or more, 80 kDa or more, 100 kDa or more, or 120 kDa or more. The composition according to claim 1 or 4, wherein the poly(vinyl alcohol) has a molecular weight of 100 kDa or less, 80 kDa or less, 50 kDa or less or 30 kDa or less. The composition according to claim 1 or 4, wherein the poly(vinyl alcohol) has a degree of hydrolysis of at least 80% or at least 90%. The composition of claim 1 or 4, wherein the poly(vinyl alcohol) has a degree of hydrolysis of less than 94%, less than 90%, less than 85% or less than 80%. The composition of claim 1 or 4, wherein each of the core particles comprises a solid crystalline drug or a salt thereof. The composition of claim 1 or 4, wherein each of the core particles comprises a solid amorphous drug or a salt thereof. The composition of claim 1 or 4, wherein each of the core particles comprises a salt of a solid medicament. The composition according to claim 1 or 4, wherein each of the core particles is a nanocrystal of a drug or a salt thereof. The composition according to claim 1 or 4, wherein the drug is at least one of a therapeutic agent or a diagnostic agent. The composition according to claim 1 or 4, wherein the drug is a small molecule. The composition according to claim 1 or 4, wherein the drug or a salt thereof has an aqueous solubility of 5 mg/mL or less, 1 mg/mL or less, or 0.1 mg/mL or less at 25°C. The method according to claim 1 or 4, wherein the drug is 80 wt% or more of the core particles, 85 wt% or more of the core particles, 90 wt% or more of the core particles, 95 wt% or more of the core particles, or 99 wt% of the core particles. The composition that constitutes the above. The composition according to claim 1 or 4, wherein the core particles have an average size of 50 nm or more or 100 nm or more and 1 μm or less. The composition according to claim 1 or 4, wherein the coated nanoparticles have an average size of 50 nm or more or 100 nm or more and 1 μm or less. 19. The composition of claim 18, wherein the average size is measured by dynamic light scattering. The composition of claim 19, wherein the average size is measured by dynamic light scattering. The composition of claim 1 or 4, wherein the coated particles have a relative velocity in mucus of greater than 0.6, greater than 0.7, greater than 0.8, greater than 0.9 or greater than 1.0. 23. The composition of claim 22, wherein the mucus is mucus of the human cervix. The composition of claim 1 or 4, wherein the composition has a polydispersity index (PDI) of 0.5 or less, 0.3 or less or 0.2 or less. The composition of claim 1 or 4, wherein the composition has a PDI of 0.1 or more and 0.5, 0.1 or more and 0.3 or 0.1 or more and 0.2 or less. 25. The composition of claim 24, wherein the PDI is measured by dynamic light scattering. 26. The composition of claim 25, wherein the PDI is measured by dynamic light scattering. The composition according to claim 1 or 4, wherein the ratio of the total weight of the drug to the total weight of poly(vinyl alcohol) contained in the composition is 1:1 to 10:1. The composition of claim 1 or 4, wherein the plurality of coated nanoparticles have poly(vinyl alcohol) present on the core particles at an average density of at least 0.01 molecules per nanometer square. The composition of claim 1 or 4, wherein the plurality of coated nanoparticles have a surface modifier present on the core particles at an average density of at least 0.1 molecules per nanometer square. The composition of claim 1 or 4, wherein the poly(vinyl alcohol) has a degree of hydrolysis of less than 95% and a molecular weight of less than 75 kDa. The composition of claim 29, wherein the poly(vinyl alcohol) has a molecular weight of 5 kDa or more, 10 kDa or more, 20 kDa or more, 50 kDa or more, 80 kDa or more, 100 kDa or more, or 120 kDa or more. The composition of claim 29, wherein the poly(vinyl alcohol) has a molecular weight of 100 kDa or less, 80 kDa or less, 50 kDa or less, or 30 kDa or less. The composition of claim 4, wherein the poly(vinyl alcohol) is PVA 13K87, PVA 31K87, PVA 9K80, PVA 2K75, PVA 57K87, PVA 105K80, PVA 130K87 or PVA 85K87. The composition of claim 4, wherein the poly(vinyl alcohol) is PVA 13K87, PVA 9K80, PVA 2K75, PVA 105K80, PVA 130K87 or PVA 85K87. The composition of claim 4, wherein the poly(vinyl alcohol) is present in the composition at 0.01% to 5% by weight. The composition of claim 4, wherein the poly(vinyl alcohol) is present in the composition at 0.1% to 5% by weight. A pharmaceutical composition comprising the composition of any one of claims 1-4 and 34-37 and at least one pharmaceutically acceptable carrier. A pharmaceutical formulation suitable for inhalation, injection or topical administration to mucosal surfaces, comprising the composition of any one of claims 1 to 4 and 34 to 37. A pharmaceutical composition comprising the composition of any one of claims 1 to 4 and 34 to 37, for delivering a medicament through the mucosal barrier. A method for forming coated nanoparticles according to any one of claims 1 to 4 and 34 to 37, comprising coating a plurality of core particles with a surface modifier to form coated nanoparticles. . 42. The method of claim 41, wherein the surface modifier is present in solution during the coating step and the agent or salt has a solubility of less than 1 mg/mL in solution at 25°C. 42. The method of claim 41 comprising simultaneously forming and coating core particles. 42. The method of claim 41 comprising forming the core particles by milling in a solution comprising a surface modifier. 42. The method of claim 41, further comprising combining the milling agent with a solution comprising core particles and a surface modifier. 46. The method of claim 45, further comprising reducing the size of the core particles with a milling agent. The method of claim 41, wherein the surface modifier is at least 0.01% (w/v), at least 0.05% (w/v), at least 0.1% (w/v), at least 0.3% (w/v) during the coating step or in the composition. , 0.5% (w/v) or higher or 1.0% (w/v) or higher in the solution.

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